Subject Index

A

• A/D converter 864

• Abbreviations 879–885

• Absorbance 842

• Acetone 246

• Acetonitrile 127, 291

• Acetonitrile gradient 459

• Acetylcholine receptor 533

• Acetylcholinesterase 676

• α₁-acid glycoprotein 490, 628

• Acid precipitation 452

• Acidic proteins 273

• Acrosin 622

• Activated charcoal 267

• Activated matrix 707

• Activated tentacle chemistry 709, 725

• Activated tentacle gels 706

• Activated thiol Sepharose 4B 645, 646

• Activation by

  • carbodiimide 719
  • cyanogen bromide 241, 621–623, 634, 719, 724
  • divinyl sulfone 621, 624
  • epichlorohydrine 724
  • epoxy 719
  • hydrazide 773
  • N-hydroxysuccinamide 719, 773
  • prohormone 655
  • tresyl 773

• Active site 460

• Additive(s)

  • see also Buffers and Detergents, Contents 839–840
  • additional substances 857–859
  • antimicrobial agents 860–861
  • buffer exchange and desalting 859–860
  • chromatographic modes

    • biospecific affinity ligands 755, 757
    • chromatofocusing 395, 404, 406
    • covalent chromatography 641, 646, 650
    • dye-ligand affinity chromatography 428–429, 445
    • hydrophobic interaction chromatography 258
    • hydroxyapatite chromatography 279, 291, 296
    • immobilized metal ion affinity chromatography 310, 320, 336–337, 339, 347, 347, 350, 375
    • integral membrane proteins 529
    • ion exchange chromatography 6, 8–9, 30, 32–33, 36–39, 48, 62, 77
    • liquid–liquid partition chromatography 480
    • reversed phase chromatography 154, 186, 199
    • scale-up of downstream processing 792–797
    • size exclusion chromatography 123, 125, 132
  • elution salts 851
  • protease inhibitors 854–857
  • thiol (SH-) reagents 854

• Additives 37, 39, 320, 337

• Adenovirus attachment protein 537

• Adenylate kinase 431

• Adenylosuccinate synthetase 431

• ADH 486, 489

• Adhesion 668

• Adhesion efficiency 685–687, 691

• Adhesion model of Hammer and Lauffenber 686

• Adhesion probability 683–684, 686

• Adhesiveness 683

• ADP 718

• 2′, 5′-ADP 306, 433, 718–719, 733, 752–759

• 2′, 5′-ADP Sepharose 755, 757

• Adrenal medulla 410

• Adsorption effects 273

• Affi-Gel 15, 622, 626, 629, 673

• Affi-Gel Heparin 745

• Affi-Prep 775

• Affinity chromatography 541, 594, 600, 603, 706

• Affinity constant 628

• Affinity displacing ligands 453

• Affinity interactions 465

• Affinity matrix 454

• Affinity partitioning 437

• Agarose 313, 672–673

• Agglutinin(s)

  • see also Lectin(s)
  • Dictyostelium discoideum 620
  • Erythrina cristagalli 633–634
  • Helix pomatia 629, 673–674, 679, 682
  • Lens culinaris 629
  • Ricinus communis 620, 629, 633–634
  • soybean lectin 684
  • Viscum album 624–625
  • wheat germ 628

• Aggregates 91, 658

• Aggregation 291, 334

• AH-Sepharose 626

• AIA 866

• Air bubbles 281

• Air degassing unit 94

• Air-free conditions 282

• Akaline phosphatase 828

• Äkta Explorer 791, 810

• Äkta 777

• D-Alanine carboxypeptidase 532

• Albumin 364, 432, 487, 490

• Alcohol dehydrogenase 113, 424, 433, 484, 486, 489–490, 492, 500, 719, 753, 759

• Aldehyde tentacle gel 714

• Aldolase 456–457

• Aldolase B 443–444, 454, 466

• Alkaline phosphatase 372, 433, 441, 827

• Alkyl chain length 532

• Alkylating reagents 641

• Alkylsulfonates 182

• Allyl-2-hydroxy-3-(N,N-dicarboxymethyl)aminopropyl ether 335

• Alpha-chymotrypsin 252

• Alpha-chymotrypsinogen 252

• Alpha-factor receptor (ste2p) (Saccharomyces cerevisiae) 537

• Alting-out effect 242

• Amberlite XAD-2 530

• D-Amino acid oxidase 430

• p-Aminophenyl glycosides 621, 626

• Amino tentacle gel 715

• 8-Aminooctyl-Sepharose 4B 293

• 8-Aminooctyl-Sepharose 352

• Aminosuccinic acid 335

• Ammonium bicarbonate 183

• Ammonium chloride 322

• Ammonium hydrogen carbonate 280

• Ammonium sulfate

  • as buffer for hydrophobic interaction chromatography 241–251, 256–267, 572, 576, 850
  • as other buffer 322, 334, 336, 405, 428, 520, 533, 570–571, 602, 620, 651, 656, 658, 739–741, 757, 799, 850, 851, 860, 862

• Ammonium sulfate precipitation 405, 602

• AMP 718

• 5′-AMP 306, 417, 718–720, 723, 733, 752–759, 851

• 5′-AMP Sepharose 757

• Amphipatic 528

• Ampholytes 8

• Ampholyte displacement chromatography 408

• Amphoteric 386

• Amphoteric molecules 387

• Amphoteric substances 849

• Amyloid P component 620

• Amylose Resin 592

• Analytical Chemistry 151

• Analytical Instrument Association 866

• Angiotensin derivative 308

• Angiotensin II 193

• Angiotensinogen 193

• Anhydrotrypsin Sepharose 572

• Animal lectins 633

• Anion exchange chromatography

  • biospecific affinity ligands 742, 750, 761
  • buffers and additives 841, 843, 845–849, 852, 858
  • chromatofocusing 387, 406
  • displacement chromatography 506, 513, 515, 519–522
  • DNA-binding proteins 596–597, 603
  • hydroxtpatite principles 272–273, 297
  • immobilized artificial membrane chromatography 459, 461, 463
  • immobilized calmodulin748
  • immobilized metal ion affinity chromatography 318, 323, 348–350, 355, 361, 363
  • ion exchange chromatography 7, 17–18, 21–23, 26, 31–33, 36, 38, 40–41, 45, 49, 52, 54–56, 59, 66, 68, 76, 79–81, 540
  • scale-up of downstream processing 772, 776, 786, 791–793, 798–799, 814, 816

• Anion exchange 515, 577, 787, 841, 843

• Anion exchanger 18, 79, 273, 461

• Anionic 845

• Anthraquinone dyes 428

• Anti-albumin 487

• Antibodies

  • see also Glycoprotein(s)
  • chromatographic modes

    • cell affinity chromatography 668, 671–673, 678, 682, 687–689, 693–696
    • chromatography on porous matrices 823
    • chromatography using Strep-tag affinity peptides 826–828
    • coupling reactions 706, 715
    • hydrophobic interaction chromatography 237–238, 257, 259, 263
    • hydroxyapatite chromatography 272, 292
    • immobilization of nucleotides 718, 721
    • immobilized metal ion affinity chromatography 310, 348, 364, 366–368, 370
    • integral membrane proteins 534, 539, 544, 547, 553, 555
    • ion exchange chromatography 9, 23, 52, 71–74, 76–78
    • lectins and glycoconjugates 620, 631, 633–634
    • liquid–liquid partition chromatography 479, 482–484, 494–498, 500–501
    • recombinant proteins 559, 562–564, 567–568
    • reversed-phase chromatography 209
    • scale-up of downstream processing 766, 768, 771, 781
  • immunoglobulins 73, 75, 119, 237, 306, 308, 335, 364–365, 369, 479, 488, 490, 568, 600–601, 633, 668

    • IgA 400, 490, 492–493
    • IgD 208, 220
    • IgE 676
    • IgG 21, 74–76, 78, 242, 315–316, 350, 479, 481, 483–484, 489–500, 544, 563, 567–568, 650, 673, 675–678, 688, 823
    • IgM 119, 238, 490, 492–493, 676
  • monoclonal antibodies 9, 52, 71–73, 78, 263, 348, 370, 394, 495, 498, 500, 539, 544, 668, 671–672, 682, 766, 823
  • polyclonal antibodies 238, 500, 534
  • screening 823

• Anti-FLAG M1 affinity gel 566

• Antigen(s) 307, 367–369, 495, 497–498, 500–501, 533, 535–537, 541, 544, 549, 559, 637, 658, 694, 823, 827–828, 830, 850

• Antigen-binding fragments 544

• Anti-IgG 487

• Antimicrobial agent(s) 201, 628, 632, 860–861

• Antipain hydrochloride 856

• Antipapain 625–626

• Antiserum 830

• Antithrombin III 734–735, 737–738, 743–745, 747

• α₁-antitrypsin 489, 490

• AP column 802

• Ap₆A 510

• apoferritin 309

• Aprotinin 856

• Aqueous polymer systems 474

• Aqueous two-phase affinity partitioning 669

• Arabinogalactan 620

• Arachidonoyl-diacylglycerol kinase 535

• Ascites fluid 365

• Ascorbate 404, 857

• Asialofetuin 627, 634–635

• Asialofetuin-sepharose 628

• Asialomucin 634

• Asialoorosomucoid 627

• Aspartase 434

• Assembly factor CsgG 827

• Asymmetry 130

• Atomic absorption spectrometry 323

• ATP 718

• ATPase 537

• ATPase complex 536

• ATP-synthase 263

• Autoclaving 743

• Autoimmune disease 497

• Automatic sample injection 283

• Automation 691

• Autoradiogram 611–613

• Autoradiography 605

• Avidin 671

• Avidin agarose 567

• Avidin-activated Biogel P-30 674

• Avidin-EDAC Biogel 689

• Azlactone 708

• Azlactone tentacle gel 715

• Azurin 828

B

• B cells 678–679, 682, 688, 693

• B lymphocytes 680

• B27 histocompatibility antigen (HLA) 537

• Bacillus stearothermophilus 429

• Bacillus subtilis 532, 693

• Back pressure 51, 112, 275, 279, 288, 296, 389, 788, 798, 806, 811

• Bacteria 767

• Bacterial protein disulfide isomerase 569

• Bacteriophage λ 588

• Bacteriorhodopsin 140, 535

• Bacteriorhodopsin fragments 538

• Bacteriophage P1 607

• Band 3 protein from erythrocyte membrane 535

• Band broadening or spreading 94, 346, 787, 789, see also Peak broadening

• Barley leaves 848

• Base matrices 775

• Basic proteins 273, 285

• Batch experiments 115

• Batch processing 874

• BBMF 706

• BCA assay 452

• Bed settling 118

• Benzyl alcohol 693

• Bestatin 856–857

• Betaine 280, 291, 320, 368, 395

• Bile acids 66

• Bile salts 530

• Bi-modal pore media 787

• Binding capacity 241, 672, 685, 750, 788

• Binding efficiency 809

• Bio–Prep SE 105

• Bio–Sil SEC 96

• Bioaffinity chromatography 541

• Bioaffinity ligands 707

• BioCAD 777, 791, 810

• BioCAD Sprint System 577

• Biocompatibility 7

• Bio-Gel A 0. 5M 673

• Bio-Gel P6 673

• Bio-Gel P30 673

• Biological activity 841, 850

• Biological buffers 845

• Biological events 845

• Biomolecules 498

• BIOSEP–SEC–S400 99

• Bio-Sil TSK series 112

• Biosilon beads 673

• Biospecific affinity gels

  • see also Contents 731–732
  • alcohol dehydrogenase 753
  • application examples 734, 739, 741, 748, 754
  • calmodulin based affinity chromatography 747
  • chromatography system set-up 737, 750
  • growth factors 733, 742
  • heparin based affinity chromatography 733–747
  • HIV reverse transcriptase 739
  • NADPH cytochrome P-450 reductase 754
  • optimization 737
  • preparing the gel 748
  • proteases 750
  • sample preparation 737, 739, 750, 755
  • serum proteins 733
  • steroid receptors 733
  • troubleshooting 760

• Biotin 671

• Biotinylated antibody 671, 678

• Biotinylated proteins 835

• Biotinylating enzyme 568

• Bis-tris 397

• Bivalent cations 404

• Bleeding

  • coupling reactions 706–707
  • IMAC 310, 312–313, 318, 323, 335, 337–338, 341, 346, 353, 360, 364–365, 376

• Blood platelet membrane 535–536

• Blood 679, 695

• Blue Dextran 681

• Blue Dye 798

• Bone marrow 695

• Borna disease virus 537

• Borosilicate glass 859

• Bound proteins 450

• Bovine IgG 74

• Bovine immunoglobulin 74

• Bovine rhodopsin 535

• Bovine serum 74

• Bovine serum albumin 56, 119, 285–286, 428, 453, 489, 603–606, 624, 671, 678, 857

• Bovine viral diarrhea virus 536

• BPG 778

• Bradykinin 193

• Brij 35 530

• BSA 487, 489, 671, 678, 798, 800, 805

• Buffer(s)

  • see Contents 839–840
  • see also Detergent(s)
  • buffers and additives 840–843, 845–851, 853, 855, 857, 859–861
  • capacities 31–33, 71, 119–120, 842
  • choice of 34–36, 38–39, 114–115, 117, 127, 243, 396, 465, 774, 844–846
  • chromatographic modes

    • biospecific affinity ligands 735, 739, 748, 750, 754, 756–757, 760–761
    • cell affinity chromatography 669, 672, 678–680, 689, 691–692
    • chromatofocusing 387, 392–396, 404–405, 408, 413
    • covalent chromatography 641, 644, 646, 649, 651, 655, 659, 662, 663
    • DNA-binding proteins 597, 602
    • dye-ligand affinity chromatography 428
    • hydrophobic interaction chromatography 238–239, 243–245, 253, 267
    • hydroxyapatite chromatography 274, 279–280, 286, 288, 290–291, 293, 296
    • immobilized artificial membrane chromatography 465
    • immobilized metal ion affinity chromatography 305, 317–321, 326, 329–330, 332, 336–337, 342–343, 346, 350, 352–356, 358–360, 364, 367, 370, 372, 374
    • integral membrane proteins 540, 543, 547, 549, 551, 553
    • ion exchange chromatography 8–9, 13–14, 33, 36, 39–41, 44, 51–52, 58–59, 62, 69, 73–74, 77, 79
    • lectins and glycoconjugates 621, 628
    • porous matrices 792–796, 798–799, 809–811, 814
    • recombinant proteins 561, 564–565, 574, 577, 582–583
    • reversed-phase chromatography 182, 186, 188–190
    • scale-up of downstream processing 766, 774, 781–782
    • size exclusion chromatography 115, 117, 123, 125–127, 132, 140
    • Strep-tag affinity peptides 830, 834
  • concentrations 123, 126, 259, 291, 392, 394, 633, 658–659, 674, 678
  • detection of 48, 245, 267
  • examples of uses 847–851
  • exchange 9, 43–44, 93, 95, 287, 859
  • flow rates 12–13, 131, 253
  • for lipids and membrane proteins 40, 42–43, 263, 464
  • ionic strengths 14, 33–35, 305
  • pH 33, 36, 52, 114, 127, 249, 394, 633, 658–659, 674–678
  • preparation of 39, 44–46, 117, 186, 245, 279, 319, 325, 329, 396, 680, 735, 748, 754, 799, 846–851
  • protein determination 245
  • removal of 287–289, 405, 859
  • selection 845–846
  • storage 132, 256, 292
  • strength 842
  • theory of 842–845
  • volatile 35–36, 280

• Buffering effect 842

• Bulk 275, 280

• Bulk gel 342

• Bulk materials 8

• 1,4-Butanediol diglycidylether coupled gel 710

C

• C1 repressor 597, 602, 607

• C1* repressor 602

• C18 alkyls 517

• Ca²⁺ ATPase 535

• Cadherin–catenin 573

• Calcium chloride 291

• Calcium phosphate 272

• Calcium-binding proteins 350

• Calibration 94

• Calibration curve 113, 130

• Calmodulin 306, 309, 318–319, 350–352, 562, 565–566, 582, 733, 747–752, 826

• Calmodulin agarose 566

• Calmodulin based affinity chromatography 747

• Calmodulin binding peptide 562, 565–566, 582, 747, 826

• Calnexin 463

• Calpains 751

• c-AMP 718

• Candida krusei 305

• Candida mycoderma 759

• Canine distemper virus 537

• Capacities 242, 275, 288, 847

• Capillary electrophoresis 78, 94, 376, 513

• Capillary electrophoresis apparatus 94

• Capillary zonal electrophoresis 814

• Caprine arthritis-encephalitis lentivirus 538

• Capture (Initial step) 272, 287, 341, 734, 813

• Carbamat bond 708

• Carbodiimide 626, 707, 719

• Carbohydrates 668, 678

• Carbohydrate microheterogeneities 217

• Carbonyldiimidazol 707, 708

• Carboxylates 273

• Carboxymethyl 7–8, 22, 26–27, 79, 304, 306, 311, 316, 335, 376, 515, 520–521, 524, 596, 772–793

• Carboxymethylated aspartic acid 335

• Carboxymethyldextran 8, 515

• Carboxymethylpicolylamine 335

• Carboxypeptidase G 433

• Cardiac membrane proteolipids 538

• Carrier 513

• Carrier fluids 774

• Catecholamine synthesis 410

• β-Catenin 573

• Cathepsins 655

• Cation exchange chromatography 114, 125, 207, 256, 657, 733, 756, 772

  • buffers and additives 843, 847, 852
  • displacement chromatography 506, 519, 520–522
  • DNA-binding proteins 596–597, 600, 603–606
  • immobilized metal ion affinity chromatography 348, 354, 361
  • ion exchange chromatography 7–9, 16–17, 21–23, 26–27, 31, 33, 36–38, 47, 53–54, 56, 59, 66, 71, 74–77, 79–81
  • porous matrices 786–787, 791, 793–794, 820, 822

• Cationic 845

• CBG 489

• Cell affinity chromatography 667–702

  • see Contents 667–668
  • application examples 694–696
  • automation 691
  • blood 679, 695
  • CAC kits 678–679
  • cell preparation 679–680
  • column testing 681
  • combinations with other chromatographies 693–694
  • gels for CAC 672, 674, 687–689
  • monoclonal antibodies 672, 682
  • non-specific cell adsorption 670
  • optional equipment 691
  • sample loading, washing, and elution 682–683
  • scaling up and down 690
  • starting procedures 679
  • stem cell isolation 695
  • troubleshooting 696

• Cell detachment 684

• Cell disruption 561

• Cell entrapment 687

• Cell function 672, 688

• Cell immunoaffinity chromatography 674

• Cell preparation 679

• Cell removal 684, 687

• Cell separation 373

• Cell surface probing 373

• Cell-CAM 105 535

• CELLector Flasks 695

• Cellulofine GH-25-c 673

• Cellulose 105, 113, 673

• Centricon 117

• Centrifugation concentration 117

• Centroid volume 138

• CEPRATE 679, 694–695

• CEPRATE Gel 673

• Ceramic hydroxyapatite 291

• Ceramic materials 275

• Ceruloplasmin 490

• Cetyltrimethylammoniumbromide 529

• cGMP (current Good Manufacturing Practice) 766–767, 782

• Channelling 131

• Chaotropic 242

• Chaotropic agents 396

• Chaotropic effect 850

• Chaotropic reagents 131

• Chaotropic salts 364, 826–827

• CHAPS 529—530, 853

• Characteristic applications 519

• Charge clusters 7

• Charged functional groups 387

• Charged groups 22

• Chelat 650 316

• Chelating buffers 291

• Chelating Sepharose 566

• Chelating Sepharose Fast Flow 314–315, 342, 370

• Chelating Sepharose High Performance 324

• Chelating Superose 315, 566

• Chelation 304

• Chelex 100 267

• Chicken breast muscle 15

• Chicken egg-white cystatin 828

• Chimeric IgG antibodies 496

• Chitin 620

• Chitin beads 565

• Chitin binding domain 562

• Chlorhexidine 861

• Chlorophyll-a/b-binding protein 349, 535

• Chloroplast ATPase 61

• Chloroplast energy coupling complex 536, 538

• Cholate 529, 853

• Cholesterol 658

• Cholesterol transfer protein 463

• Choline acetyl transferase 432

• Chromaffin granules 410

• Chromaflow 778

• Chromatofocusing

  • see Contents 385–386
  • application examples 408–413
  • choice of gel 389, 396
  • column packing 400
  • determination of isoelectric points 406–407
  • ion exchange chromotography 624
  • marker proteins 400
  • Mono P 390–391, 409–410
  • optimal conditions 403–404
  • pH-gradients 387
  • principle 387–388
  • sample preparation 398
  • scaling up 406
  • set-up 396–397
  • test separation 400–402
  • troubleshooting 258, 413
• Chromatography software 863–876

  • see Contents 863
  • basic necessities 865–866
  • commercially-available 868
  • HPLC software packages 875
  • interesting features 867
  • package examples 865
  • support 875
  • training courses 875

• Chromophore 70

• CH-Sepharose 626

• Chymase activities 355

• Chymosin 430

• Chymostatin 625–626, 856

• Chymotrypsinogen 244, 247

• α-Chymotrypsin 78

• α-Chymotrypsinoge 285

• α-chymotrypsinogen A 286

• Cibacron blue F3G-A 416

• CIP

  • see Cleaning in place

• Circular dichroism spectrometer 94

• Circular dichroism 140, 142

• Citrate 289

• Cleaning in place 150, 256, 652, 670, 693, 706, 781

• Clostridium perfringens 680

• Cloud point 66, 532–533

• Cloudy protein solutions 280, 395

• CM 7

• CMC 64, 123, 529

• CMC

  • see Critical micellar concentration

• CMP 718

• CM-Sephadex 656–657

• CNBr 720, 724

• CNBr-activated sepharose 572

• CNBr-activated Sepharose 6MB 689

• Coagulation factors 306

• Coagulation proteins 734

• Code system 620

• Collagenases 306

• Colostrum 726

• Column bed height 776

• Column bed volumes 780

• Column calibration 93

• Column conditioning 187

• Column cross – sectional area 799

• Column diameter 129, 790

• Column dimensions 780, 808

• Column library 872

• Column lifetime 191

• Column log-book 811

• Column oven 93

• Column packing 800

• Column size 808

• Column switching 319

• Column(s)

  • see also Gel(s)
  • antimicrobial agents
  • calibration 89, 91, 93–94, 113, 115, 117–118, 127–128, 130, 133, 143, 188, 239, 246, 326–328, 481, 621, 624, 738, 866
  • dimension(s) and bed volume(s) 10–11, 15, 25, 42, 46–49, 55–59, 73, 91, 122, 190, 128, 246, 248, 253–256, 260, 262, 264, 282, 285, 292, 320–344, 365–366, 370, 375, 391, 394, 397–400, 403, 406, 409, 413, 439, 474, 479, 486, 508, 517–518, 566, 575, 624–628, 634, 647, 658, 672, 677–678, 680–683, 690–691, 734, 771, 775–781, 790, 798–802, 806–809, 813, 817–821, 831, 833
  • empty 10, 46–47, 280–281, 342–343, 399, 408, 776–779, 800, 802–803
  • packing of

    • cell affinity chromatography 680–681
    • chromatofocusing 399–400
    • covalent chromatography 646, 648–649
    • hydrophobic interaction chromatography 245–246
    • hydroxyapatite chromatography 276, 280–281
    • ion exchange chromatography 46–47
    • reversed-phase chromatography 156, 157, 176, 194
    • size exclusion chromatography 118
  • size 11, 117, 128, 244, 251, 259, 261, 290, 336, 404, 445, 459, 486, 517, 776, 781, 808
  • stabilization 38–41, 95, 622, 754

• Column-ovens 155

• Columns in tandem 319

• Commercially available 437

• Compaction 687, 689

• Compartmentation 841

• Compatibility 864

• Competitive ligand 321

• Competitive ligand strength 322

• Complex protein samples 409

• Compression 131, 275

• Computer 10, 11, 15, 62, 409, 451, 791, 810

  • see also Software chromatography software 863–876

• Concanavalin 684

• Concanavalin A 620

• Concanavalin A tentacle affinity matrix 714

• Concentration 847

• Conductivity detector 513

• Conformational changes 500

• Conformational structure 237

• Connecting a ligand 724–725

• Contact time 683

• Contain peroxides 851

• Continuous bed chromatography 79

• Convective mass transfer 789

• Convective mass transport 787

• Corona virus 533

• Corticosteroid-binding globulin 490

• Cost(s)

  • biospecific affinity ligands 742
  • buffers and additives 854
  • cell affinity chromatography 668–670, 687, 689, 693
  • chromatofocusing 408
  • chromatography on porous matrices 788, 814
  • chromatography software 867
  • coupling reactions 706
  • covalent chromatography 650
  • dye-ligand affinity chromatography 440
  • hydrophobic interaction chromatography 238, 251
  • hydroxyapatite chromatography 275, 290
  • immobilized metal ion affinity chromatography 310, 314, 342
  • integral membrane proteins 529, 533–534
  • ion exchange chromatography 24, 55
  • recombinant proteins 564, 568
  • reversed-phase chromatography 194
  • scale-up of downstream processing 771, 774, 782
  • software systems
  • size exclusion chromatography 116, 124, 133, 140, 143

• Coulter counters 671

• Coupled buffer 289

• Coupling reactions

  • see Contents 705
  • bleeding 706–707
  • coupling reactions 706–709, 712–715
  • spacer technologies 709
  • synthesis examples 712–715
  • tentacle gels 711–712, 714–715

• Coupling to grafted polymer chains 709

• Coupling to spacers 709

• Covalent chromatography

  • see Contents 639
  • additives
  • application examples 694–696
  • direct and analytical modes 646, 648
  • DTT 654
  • excess active groups 648
  • gels commercially available 645
  • intramolecular disulfide bonds 641
  • 2-mercaptoethanol 641
  • optimization 651–652
  • scale-up 652
  • Triton X-100 652
  • troubleshooting 662–663

• Creatine kinase 433

• Critical micellar concentration 52, 63–65, 115–116, 123, 529–532, 541, 548, 852

• Cross-linked agarose 95

• Crosslinked polyacrylamide 313

• Crosslinked polysaccharides 94

• Crosslinked polystyrene 95

• Crotalus atrox 458, 460–461, 466

• Crude egg white 409

• Crude extracts 150, 237, 260, 347, 442, 630, 857

• Crystallization attempts 141

• Crystallization 93, 620, 812

• CTAB 65, 852–853

• CTP 718

• Cuatrecasas 724

• Current Contents 151

• C_xE_y series 531

• Cyanobacteria 814

• Cyanobacterial phytochrome 828

• Cyanobacterium 295, 349

• Cyanogen bromide 239, 241, 572, 595, 597, 600, 621–623, 626–627, 634, 640, 645–646, 659, 672–677, 679, 689, 696, 707–708, 719–721, 724, 744, 747, 748–749, 757, 759, 772

• L-Cysteine agarose 645

• Cyt b ₆ /f and LCHII 537

• Cytochrome b₅ 7, 294, 462–463, 563, 567–568

• Cytochrome b-561 410

• Cytochrome b ₅₆₂ 828

• Cytochrome b ₆ f 294

• Cytochrome b ₆ f complex (synechocystis PCC 6803) 535

• Cytochrome c 78, 193, 244, 247, 249, 252, 285, 286, 392, 400–401

• Cytochrome(s) c

  • chromatofocusing 392, 400–401, 411, 413
  • chromatography using Strep-tag
  • affinity peptides 827–828
  • hydrophobic interaction
  • chromatography 244, 246–247, 249–252
  • hydroxyapatite chromatography 285–286
  • immobilized artificial membrane chromatography 462
  • immobilized metal ion affinity chromatography 304–306, 309, 319, 324–326, 330, 332–333, 335, 339, 376
  • integral membrane proteins 535–538
  • ion exchange chromatography 67, 78
  • recombinant proteins 558, 583
  • size exclusion chromatography 113, 119

• Cytochrome c oxidase 535, 558, 827, 833

• Cytochrome c oxidase (Bacillus PS3) 536

• Cytochrome c oxidase subunits 536, 538

• Cytochrome c reductase (potato

• mitochondria) (Neurospora crassa) 537

• Cytochrome c reduction 462

• Cytochrome(s) P-450

  • biospecific affinity ligands 754, 756, 759
  • hydroxyapatite chromatography 278, 287–289, 293–294
  • immobilized artificial membrane chromatography 451, 453, 461–463, 466
  • immobilized metal ion affinity chromatography 323, 352–355
  • integral membrane proteins 536, 538, 547–548
  • ion exchange chromatography 13, 23, 33, 36, 41–43, 48–51, 54–55, 57, 65–67

• Cytochrome P-450 fragments 538

• Cytochromes (E. coli) 535

• Cytolomegalovirus 498

• Cytolytic granules 355

• Cytomegalovirus 537

• Cytosolic proteins 237

D

• Damköhler number 685

• Data acquisition 873

• Data buffer buffered interface 865

• Data-bases for file organization 873

• DDE 866

• DDT 605

• De novo synthesis 138

• DE52 cellulose 569, 572

• Dead space 131

• DEAE 7–8, 18–20, 22, 25–26, 29–30, 40–41, 45, 47, 50–52, 56, 60, 67–68, 223, 258, 308, 350, 361, 370–371, 418–419, 421, 423, 441–442, 444, 463, 481, 511, 519–521, 596, 598–599, 631–632, 646, 649, 654, 656–657, 662, 759, 772, 792–793

• DEAE-cellulose 632

• DEAE-dextran 8

• DEAE-Sephacel 258

• Degassing 93, 117, 279, 392, 409

• Degradation products 272

• Degree of dye substitution 436

• Dehydrofolate reductase 661

• Dehydrogenases 718

• Delipidation 336

• Denaturation 291

• Denaturing agents 314

• Density gradient centrifugation 680

• Deoxycholate 65–66, 121, 140–141, 452, 529–530, 534, 537, 629, 633, 853

• Depth filter 685

• Depth filtration 685

• Depyrogenated 692

• Depyrogenating 693

• Desalting 90, 95, 104–105, 150–151, 183, 217, 257, 323, 336, 338, 340, 348, 438, 512, 589, 597, 604–605, 646, 648, 650, 656, 660, 662, 742, 760, 846, 850, 857–860

• Desalting column 340

• Desalting in reversed-phase 217

• Desthiobiotin 831

• Detection 682, 791, 804, 865

• Detergent(s)

  • additives 291
  • chromatographic modes

    • affinity chromatography 541–545, 683–689, 751, 759
    • hydrophobic interaction
    • chromatography 237–264
    • hydroxyapatite chromatography 273–289
    • immobilized artificial membrane chromatography 452–465
    • immobilized metal ion affinity chromatography 314–344
    • ion exchange chromatography 9–68, 540–541
    • reversed-phase chromatography 217–223, 545–547
    • size exclusion chromatography 92–141, 537–540
  • cloud point 532–533
  • compatibility with eluents 66, 652, 804, 852–854
  • compatibility with gels 67, 249, 320, 337–338, 452–454, 652, 683, 689
  • concentrations 123–124, 537, 683, 751, 817, 834, 852–854
  • exchange 117, 140
  • flow rates 131, 804, 814, 816, 834
  • general 9, 23, 92, 115, 237–238, 273, 280, 314, 364, 457, 462, 528, 841, 852
  • interference 454, 529, 561, 622, 629, 751, 814, 816, 852–854
  • lectins 622–634
  • membrane proteins 61–68, 115–117, 120–121, 123–124, 281, 528–547, 622, 751, 814, 816, 852–854
  • microsomes 43, 48, 759
  • molecular mass 118, 134, 136, 141, 529–530, 532
  • porous matrices 804–834
  • properties 61–64, 278, 529–534, 852–854
  • protein structure or function 65–66, 120
  • recombinant proteins 561–579
  • reference reviews 68, 529
  • removal of detergent

    • amberlite incubation 530
    • comparison of gel types 289
    • by dialysis 529
    • efficiency 141
    • eluent properties 287–288
    • examples 68, 287–288
    • by hydrophobic interaction chromatography for non-ionics 237
    • by hydroxyapatite chromatography for non-ionics from membrane proteins 273
    • important detergent properties 532
    • by ion exchange chromatography 68, 141
    • matrix type and pH 58
    • reference reviews 68
    • regeneration with ethanol 58
    • prior to reversed-phase chromatography 217, 223
    • by size exclusion chromatography 141, 529
    • wavelengths 67, 339
  • resolution by 66–67
  • selection of 33–34, 37–39, 41, 61–64, 115–116, 465, 529, 652
  • solubility efficiency 64–65, 123–124, 131, 249, 264, 337–338, 465, 561, 579, 622, 629, 652, 683, 751, 759, 814, 816, 827, 834, 852–854
  • temperature 51, 66, 118, 344, 561, 827, 852–854
  • wavelength range 67, 116, 287, 339–340, 533, 852–854

• Detergent step gradient 460

• Detergent-solubilized DMPE 453

• Dextran 118, 471, 620, 672–673, 706

• Dextran 40 480

• Dextran 500 480

• Dextransulfate 8

• Diabetes mellitus 70

• Diafiltration 767

• Dialysis 117, 529, 860

• Diamid bond 708

• Diaminobiotin 565, 827

• Diazepam 193

• Dichloromethane 132

• Dichlorotriazine dyes 436

• Dictyostelium discoideum 620

• Diethanolamine 402

• Diethylaminoethyl 7

• Differential refractometer 137

• Diffusion paths 787

• Diffusion pores 787

• Diffusion-controlled process 93

• Digitonin 852–853

• Dihydrodiol dehydrogenase 660

• Dihydrofolate reductase 308, 358–360, 362, 431, 661, 663

• Dihydrogenphosphate 279

• Dimerization 118

• Dimethyl sulfoxide 190, 321, 859

• Dinucleotide binding fold 416

• Diol tentacle gel 712

• Diol 95

• Dipeptidyl aminopeptidase 535

• Diphenhydramine 193

• Dipicolylamine 335, 363

• Diptheria toxin 431

• 2, 2′-Dipyridyl-disulfide 642

• Direct coupling 709

• Directory of manufacturers and suppliers 887–894

• Displacement chromatography

  • see Contents 505
  • application examples 522
  • characteristic applications 519
  • column size 517
  • elution modes in 506
  • gels in 508
  • ion exchange chromatography 79
  • location in a purification scheme 508
  • MALDI-MS 513
  • principle 506, 508
  • system set-up 512
  • troubleshooting 522

• Displacement train 509

• Displacer molecules 8

• Displacing salts 38

• Distribution coefficient 91

• Disulfide bonds 141, 571, 641

• Disulfide bridge 625

• Dithiothreitol 37, 39–40, 43–45, 243, 261, 289, 291, 294, 321, 334, 407, 409, 431–432, 435, 545–546, 576, 576, 602, 604, 607–609, 641–642, 648–650, 652, 654–660, 663, 754–755, 841, 854

• Divinyl sulfone 621–622, 624

• Divinyl sulfone-activated Sepharose 634

• DMSO 190, 265, 859

• DNA 307

• DNA polymerases 739

• DNA-binding proteins

  • see Contents 587
  • ammonium sulfate precipitation 602
  • application examples 607–614
  • autoradiography 605, 611–613
  • commercially-available gels 598, 600
  • desalting 604
  • DNA affinity chromatography 595
  • DNA mobility shift assay 609
  • DNA-binding proteins 587–595, 607–609, 611–613
  • DNase I footprint 611
  • Heparin Sepharose 597, 589, 602–603, 605
  • Klenow fragment 609, 611–613
  • operator 588
  • protein-DNA interactions 587
  • purification tag 590
  • specific site 606

• DNase I footprint 611

• DNAse 812

• DOC

  • see Deoxycholate

• Dodecyl-β-D-maltoside 296, 530

• β-Dodecylmaltoside 814

• Dodecyl-maltopyranosid 834

• Domain-specific binding 7

• Donnan potential 114, 406

• Dot polymer 711

• Downstream processing 706, 826

• DP 711

• Drivers 866

• Dry powder 281

• DTE 40, 321

  • see Dithioerythritol
• DTT 40, 289, 291, 321, 654

  • see Dithiothreitol

• Dual-wavelength scanner 452

• Duffy protein 658

• Dye affinity chromatography 707

• Dye leakage 428

• Dye-ligand affinity chromatography

  • see Contents 415
  • application examples 441–444
  • column preparation 438–439
  • column regeneration and storage 440
  • commercially-available dye-liganded gels 418–426
  • coupling of reactive dyes 429
  • dye leakage 428
  • dye-ligand affinity chromatography 416, 429–437, 441–444
  • dye-ligand screening kits 437
  • operational parameters 429
  • protein purification examples 430–435
  • reactive dye chemistry 428
  • sample preparation 437
  • screening of dye-ligands 442–443
  • spacer molecules 417
  • stacking effects 428
  • troubleshooting 444–445

• Dynabeads 373, 695

• Dynamic data exchange 866

E

• E. coli

  • chromatography using Strep-tag affinity peptides
  • DNA-binding proteins 589, 593–595, 597, 608–609
  • dye-ligand affinity chromatography 432, 434
  • hydroxyapatite chromatography 294
  • immobilized metal ion affinity chromatography 360–362, 367, 372
  • integral membrane proteins 535–538
  • ion exchange chromatography 40
  • miscellaneous biospecific affinity ligands 739, 741, 747–748, 754–756
  • recombinant proteins 559–562, 565, 568–569, 571, 573–575, 580
  • reversed-phase chromatography 215–216
  • size exclusion chromatography 136

• E1 glycoprotein 533

• E-64 856

• Eddy diffusion 80

• EDTA 38, 291, 322, 404, 605, 654, 856–857

• Effectively shadowing 685

• Efficiency 112

• Efficiency-term 128

• Egg white 409

• EGTA 322

• Electron crystallography 814

• Electrophoretic separation 386

• Electrostatic effect 406

• ELISA 529

• Eluent 393–394

• Eluotropic mobile phase 453–454

• Eluotropic strength 452, 458, 464–465, 546

• Eluting agents 321

• Elution 827

• Elution buffer 394

• Elution buffer strength 353

• Elution chromatography 506, 511

• Elution modes 506

• Elution salts 851

• Elution volume 91

• Emulgen 273, 287

• Emulgen 911 43, 287, 293, 462

• Emulphogen 530

• Endcapped 174, 461

• Endopeptidases 855

• Endoplasmic reticulum 461

• β-Endorphin 747

• Enterokinase 563

• Entrapment of cells 672

• Enzymatic digestion 812

• Enzymatic lysis 561

• Enzyme immobilization 640

• Epichlorohydrine 620, 724

• Epichlorohydrin-coupled gel 710

• Epidermal growth factor 537, 542

• Epidermal growth factor receptor 537

• Epoxide 292, 798

• Epoxy 719

• Epoxy-activated gel 710

• Epstein–Barr virus 536–537

• Equilibration buffer 465

• Equilibrium binding capacity 32

• Equilibrium constant 137

• Equine infectious anemia virus (EIAV) 535

• Erythrina cristagalli agglutinin 633, 634

• Erythrocyte ghosts 535

• Erythrocyte(s) 373, 431, 434, 535–537, 623, 631, 658, 664, 676, 681–682, 692

• Erythrocytes (human) (e.g. glycophorin C) 535–537

• Escherichia coli 40, 826

• Escherichia coli overproducer 589

• Ester linkages 460

• ^esteriam.PC 451, 462–463, 466

• Ethanol 291, 861

• Ethanolamine 402, 848

• ^etherIAM.PC^Cl0/C3 451, 454, 456–460, 466

• ^etherIAM.PE^Cl0/C3 451, 453, 455, 457

• ^etherIAM.PS^C10/C3 451, 457

• Ethylation footprint 613

• Ethylene glycol 249, 291, 321, 628

• Ethylene oxide 692

• Eucaryotes 767

• Eucaryotic cells 373

• Exopeptidases 855

• Expression systems 559

• Expression vector 591–593

• Extracellular matrix 733

• Extraction 531

F

• F1-ATPase 433

• F₁F₀-ATPase (bovine heart mitochondria) 536

• Fab fragments 76

• Fab 495

• FACS 695

• Factionation by LLPC 492

• Factor IX concentrate 369

• Factor Xa 563

• Fast desalting 105

• Fc portion 366

• FDH 489

• Ferric adsorbent paper 373

• Ferric ions 499

• Ferritin 309

• Ferritin 486

• Fetal calf serum 678

• Fetuin 628

• Fibrinogen 306

• Fibroblast growth factor 308

• Fibronectin 306, 309, 622

• Ficoll sedimentation 695

• Field flow fractionation 669

• Filter membranes 279, 395

• Filter 117

• Filtration 186, 216, 767

• Final polishing 272

• FineLINE 778

• Fines 281

• First separation 284

• FITC 688

• FITC-conjugated avidin 671

• FLAG 560

• FLAG-peptide 562, 564

• Flavanone 3 β-hydroxylase 407–408

• Flow adapter 670

• Flow chambers 684

• Flow cytometers 671

• Flow rate 20, 37, 57, 78, 123, 128, 157, 162, 164, 166, 170, 175, 177, 179, 181, 193, 248, 255, 257, 276, 328, 331, 338, 364–366, 391, 452, 463, 465, 478, 485, 498, 502, 510–511, 516, 519–520, 736, 788–790, 799, 805, 820, 833, 846

• Flow-chart 867, 873

• Flower color 294, 408

• Flow-through system 685

• Fluorescence 122

• Fluorescence microscopy 682

• Fluorescent activated cell sorters 668, 671

• Fluorescent activated cell sorting 669

• Fluorophore-conjugated antibody 671

• FMOC 819

• Focused polypeptides 388

• Focusing effect 388

• Folded protein 560

• Folding 142–143

• Formaldehyde 693

• Forssman glycolipid 629

• Four-helix dimer 138

• FPLC (fast protein liquid chromatography)

  • affinity chromatography 740
  • buffers and additives 860
  • chromatofocusing 389, 409–410
  • chromatography on porous matrices 789, 798
  • chromatography software 865–868
  • DNA-binding proteins 595, 597
  • hydrophobic interaction chromatography 239, 244
  • hydroxyapatite chromatography 274, 276
  • immobilized metal ion affinity chromatography 311, 313–314, 321, 324–325, 327–330, 338, 341, 343, 345–346, 352–353, 355, 371
  • ion exchange chromatography 10–11, 13, 15, 25–26, 44, 55, 58–59, 71–72
  • recombinant proteins 564, 569, 572

• FPLC 274, 389

• FPLC system 327, 409

• Fraction collection 11, 13–14, 16, 48, 74, 122, 155, 247, 259, 263, 274, 311, 324, 339, 347, 389, 401, 472, 518, 569, 577, 597, 619, 647, 691, 734, 737, 748, 750, 754, 791, 804, 810, 863, 866, 869

• Fraction collector 14, 791

• Fractogel EMD Azlactone 650 (S) 713

• Fractogel EMD Chelat 650 (M) 713

• Fractogel EMD Chelat 650 (S) 713

• Fractogel EMD Epoxy 650 (S) 713

• Fractogel EMD TA 650 (S) 713

• Fractogel EMD 105, 316

• Fractogel 775

• French Press 831

• Frontal boundary analysis 137

• Frontal chromatography 506

• Fucan 632

• Fucoidan-Sepharose 630, 632

• Fumarate reductase 430

• Fusion partners 560

• Fusion protein 306–308, 357, 590

• Fusion protein cleavage 591–593

• Fusion 591–593

• Fusogenic protein (rat brain microsomal membranes) 537

• F_v fragment 367, 833

G

• G protein 533

• G1000PW 109

• G300SW_XL 99

• GAG 746

• Galactosamine 679

• Galactose-specific proteins 621

• β-Galactosidase 309, 563

• Galactoside-binding lectin 624

• Galactoside-binding proteins 635

• β-Galactoside-binding proteins^a 627, 633

• Galactoside-specific lectin 625

• Galactosyl-specific antibodies 620

• Galaptin 633

• Galectin-1 625

• Galectins 622, 633

• Gas chromatography 868–873

• Gastric mucin 621

• GC200 105

• GDH 489

• GDP 718

• Gel filtration chromatography

  • see Size exclusion chromatography

• Gel filtration experiments 853

• Gel filtration 860

• Gel matrices 564

• Gel permeation chromatography 91

• Gel permeation chromatography

  • see Size exclusion chromatography

• Gel permeation 656

• Gel suspension 281

• Gel(s)

  • see also Column(s) and Stationary phase(s)

    • capacity

      • cell affinity chromatography 671–673, 685–692, 697
    • chromatofocusing 391–392, 394, 398, 403–405
    • chromatography on porous matrices 787, 788, 791–792, 794, 796, 798, 806–813, 819–820
    • chromatography using Strep-tag affinity peptides 831, 835
    • coupling reactions 706, 709, 713
    • covalent chromatography 644–645, 648, 651, 654, 660–662
    • displacement chromatography 507, 513, 515
    • DNA-binding proteins 594
    • dye-ligand affinity chromatography 417–426, 436, 438
    • hydrophobic interaction chromatography 239, 241–242, 252–254, 256
    • hydroxyapatite chromatography 272, 275–276, 278–279, 290
    • immobilized artificial membrane chromatography 464
    • immobilized metal ion affinity chromatography 312–318, 334, 336, 341–343, 346, 350–353, 263–366, 373, 375
    • ion exchange chromatography 9–11, 17, 19–20, 24, 26–27, 31–33, 35–36, 42, 54–56, 59, 66, 69, 71, 76–77
    • lectins and glycoconjugates 621–622, 626, 628–630, 634–635
    • liquid–liquid partition chromatography 475, 481–482
    • miscellaneous biospecific affinity ligands 733–734, 738, 743–744, 747, 749–750, 752, 757–758
    • recombinant proteins 566–568
    • reversed-phase chromatography 152, 157, 160, 162, 164, 168, 170, 176–177, 179, 188–189
    • scale-up of downstream processing 774, 776, 781
    • size exclusion chromatography 92, 122, 127–129
  • commercially-available

    • 2′, 5′-ADP and 5 ′-AMP Sepharose 757
    • anion exchange chromatography 18–20
    • calmodulin Sepharose 749
    • cation exchange chromatography 21–22
    • cell affinity chromatography 673
    • chromatofocusing 390
    • chromatography on porous matrices 792–797
    • coupling reactions 713
    • covalent chromatography 645
    • DNA-binding proteins 598–601
    • downstream processing for large scale 772–773, 775
    • downstream processing or technical scale 778–779
    • dye-ligand affinity chromatography 418–425
    • heparin affinity chromatographic media 744–745
    • hydrophobic interaction chromatography 240
    • hydroxyapatite chromatography 276–277
    • immobilized metal ion affinity chromatography 315–316, 335
    • liquid-liquid partition chromatography 473
    • recombinant proteins 562–563, 566–567
    • reversed-phase chromatography 157–173
    • size exclusion chromatography 96–111
  • equilibration 118, 121, 326, 329, 332, 439–440, 443, 452, 464–465, 480–481, 508, 540, 569, 597, 650, 680, 750, 755, 801

    • chromatofocusing 392–394, 397, 400–403, 409–410, 413
    • hydrophobic interaction chromatography 245–246, 250, 259–260, 262, 264
    • hydroxyapatite chromatography 273, 277, 279, 282, 284–287, 293–294
    • ion exchange chromatography 9, 39, 41, 44–49, 69, 71, 73, 75
    • reversed-phase chromatography 152, 180, 187, 190, 192
• regeneration 841, 848

  • • cell affinity chromatography 671, 683, 689, 693
    • chromatofocusing 402–403, 413
    • chromatography using Strep-tag affinity peptides 831, 833
    • dye-ligand affinity chromatography 440
    • hydrophobic interaction chromatography 238, 256–257
    • hydroxyapatite chromatography 266–277, 291–292
    • immobilized artificial membrane chromatography 453
    • immobilized metal ion affinity chromatography 306, 313–314, 321, 324–325, 330, 332, 334, 345, 346
    • ion exchange chromatography 9, 29, 49, 58–59, 66, 71
    • lectins and glycoconjugates 628–629, 652
    • liquid–liquid partition chromatography 488, 509, 512–513
    • miscellaneous biospecific affinity ligands 743, 748, 751, 758
    • reversed-phase chromatography 189–190
    • scale-up of downstream processing 771, 774, 811
    • size exclusion chromatography 131–132
  • stabilization 38–41, 95, 622, 754
  • storage 59–60, 132–133, 191, 256, 291–292, 390, 403, 440, 451, 488, 577, 628–629, 652, 741, 743, 751, 758, 774, 812, 853–858
  • washing

    • cell affinity chromatography 679, 681–684, 690–691, 693, 695
    • chromatofocusing 402, 409, 411–412
    • chromatography on porous matrices 800–801, 804, 806, 808, 811–813, 827, 829, 831, 834
    • coupling reactions 706, 716
    • covalent chromatography 643–644, 646, 651–652
    • dye-ligand affinity chromatography 428, 439–440, 444
    • hydrophobic interaction chromatography 243, 247, 256, 259–260, 263–266
    • hydroxyapatite chromatography 287–288, 291–293, 304, 311, 321, 323, 326, 329, 331, 333, 338, 341, 344–345, 352, 368–370, 375
    • immobilized artificial membrane chromatography 453–454, 456, 474
    • integral membrane proteins 544
    • ion exchange chromatography 13, 44, 4f3–49, 57–58, 60, 69–70, 73
    • miscellaneous biospecific affinity ligands 751, 755, 758, 761
    • recombinant proteins 570–572, 576–577, 579, 621, 626, 628, 634
    • reversed-phase chromatography 187, 190–191, 197
    • size exclusion chromatography 112, 135

• Gelfiltration 405

• Generic expression vectors 833

• GFC (Gel filtration chromatography)

  • see Size exclusion chromatography

• GFP 831

• ^δGIAM.PC^C10/C3 451, 460, 454

• Glass 673

• Glass beads 672, 679

• Glass columns 670

• Glass matrices 314

• Glossary for ion exchange 79–81

• GLP (Good Laboratory Practice) 867, 869–873, 875

• Glucocerebrosidase (human placenta) 535

• Glucokinase 435

• Glucose 6-phosphate dehydrogenase 435, 754

• Glucose oxidase 72

• Glucose transporter protein 463

• Glucose transporter 535

• Glucose-DH 486

• Glutamate dehydrogenase 719

• Glutaraldehyde 622

• Glutathione 571

• Glutathione agarose 567

• Glutathione reductase 719

• Glutathione Sepharose 590–591

• Glutathione-S-transferase 563, 565–566, 659–660

• Glutathione transferase 308

• Glycerol backbone 460

• Glycerol 404, 858–859

• Glycerylpropyl bonded surface 112

• Glycinehydroxamate 335

• Glycoconjugate 625, 726, 713

  • see also Lectins and glycoconjugates

• Glycohistochemical monitoring 632

• Glycol 471

• Glycoligand 620–621, 630

• Glycolipid 629

• Glycophorin 373

• Glycoprotein D 544

• Glycoprotein(s)

  • see also Lectins and glycoconjugates
  • ion exchange 40, 67, 69–70, 72
  • membrane proteins 533, 535, 541, 544
  • reversed-phase 202, 217, 220–221

• Glycosaminoglycan 746–747

• Glycosidase 620, 623, 634

• Glycosylation 195

• Glycosylation patterns 632

• Glycosyltransferase 620, 634

• Glycyl tyrosine 130

• GMP (Good Manufacturing Practice) 706, 718, 726, 875

• GMPWXL 111

• Goldstein 620

• Gonadotropic hormones 306

• Good laboratory practice 867

• Good-buffer 845, 847

• Gossypium hirsutum 463

• GPC (Gel permeation chromatography)

  • see Size exclusion chromatography

• Gradient anomalies 737

• Gradient conditions 465

• Gradient detergent elution 453

• Gradient elution 80

• Gradient mixer 13

• Gradient mobile 452

• Gradient salt elution 453

• Grafted-tentacle gel 711

• Granulocytes 672

• Granzymes 354

• Grapevine leaf-roll associated virus 859

• Gravity flow 670

• Green fluorescent protein 831

• Growth factors 306, 733, 742

• Guanidine hydrochloride 743, 835, 859

• Guanidine 131, 141, 292, 314

• Guanidinium chloride 38

• Guanidinium hydrochloride 127

• Guard columns 452

• Guideline for binding and elution 286

H

• HA or HAC

  • see Hydroxyapatite chromatography

• HABA 831

• Haemacytometer 671

• Haemagglutination assays 623, 630

• Haemolysin 658

• Halobacterium halobium 140, 535

• Halogen containing compounds 114

• Halophilic bacteria 9

• Handbook templates 872

• Hanks balanced salt solution 678

• Haptens 498

• Hard metal ions 350

• Hardware 864

• HA-Ultrogel 293

• HBGFs 742

• Hecameg 530

• Helix pomatia agglutinin 629, 673–674, 679, 682

• Helix pomatia-Sepharose 689

• Heme 70

• Heme groups 791

• Hemiacetal hydroxyl group 621

• Hemoglobin 70

• Henderson and Hasselbalch 842

• Heparin 798

• Heparin based affinity chromatography 733–747

• Heparin-binding growth factor 742

• Heparin-binding lectin 623, 625

• Heparin HyperD M 744

• Heparin Sepharose 589, 597, 602–603, 605, 623, 744

• Hepatic triglyceride lipase 734

• Hepatitis B 537

• Heptafluorobutyric acid 182

• Herpes simplex virus 535–538, 544

• Heterobifunctional crosslinking reagents 641

• Hexokinase 430

• HFBA 182

• Hibitane digluconate 693

• HIC

  • see Hydrophobic interaction chromatography

• (His)₆-tag 592

• His₆; His₁₀ 562

• His-bind 566

• His-patches 564

• His-tagged proteins 307, 357, 564

• Histamine 672

• Histidine 392

• Histidine residues 309

• Histones 202, 392

• HiTrap Chelating 315

• HiTrap columns 313

• HiTrap Desalting 104

• HiTrap Heparin 744

• HIV reverse transcriptase 739

• HLB values 64

• HLB 529

  • see Hydrophilic–lipophilic balance

• HMG-box 575

• Hofmeister series 248, 850

• Hollow fiber membranes 691

• Horse 304

• Hotlines 875

• HpA-Biogel P-30 687

• HpA Sepharose System 689

• HpA-Sepharose 6MB 681

• HPLC software packages 875

• HPLC

  • affinity chromatography 537, 541–545, 547
  • buffers and additives 860
  • chromatofocusing 389
  • chromatography on porous matrices 789–791, 799–804, 806, 810, 813–814, 818
  • chromatography software 864–865, 868–872, 874–875
  • displacement chromatography 507, 511–514, 518, 528
  • dye-ligand affinity chromatography 428
  • hydrophobic interaction chromatography 238–239, 244, 247, 259
  • hydroxyapatite chromatography 274, 276, 278, 534
  • immobilized artificial membrane chromatography 451, 463–464, 466
  • immobilized metal ion affinity chromatography 311, 313–314, 321, 324–328, 341–344, 352, 365, 372
  • integral membrane proteins 528, 534–549
  • ion exchange chromatography 11, 12, 15, 25–26, 39, 44, 51–52, 55, 58–59, 70, 74, 76, 79–80, 536, 540–541, 547
  • liquid–liquid partition chromatography 472, 481, 489–490
  • recombinant proteins 564, 569
  • reversed-phase chromatography 150–151, 153–155, 178–179, 185–187, 189, 191–192, 194–195, 538, 541–545–547
  • scale-up of downstream processing 766, 774, 777
  • size exclusion chromatography 91–95, 115–118, 120, 122–127, 129–130, 132–134, 136–137, 140–143, 535, 537–539, 547

• HPLC-grade water 186

• HPSEC 91

• HR Column 803

• HSA 489, 678

• Human lung leukotriene C₄ synthase 537

• Humanized murine IgG₁ 365

• Hybrid ions 291

• Hybridoma technology 766

• Hydrazine 798

• Hydrodynamic diameters 90

• Hydrodynamic instability 849

• Hydrodynamic viscosity radius 27

• Hydrodynamic volume 91

• Hydrogenphosphate 843

• Hydrophathy plots 252

• Hydrophilic heads 528

• Hydrophilic vinyl–polymer 108

• Hydrophilic–lipophilic balance number 531

• Hydrophilicity 531

• Hydrophilic–lipophilic balance 63–64, 529, 531–532, 548–549

• Hydrophobic adsorption chromatography 236

• Hydrophobic amino acid 236

• Hydrophobic domains 852

• Hydrophobic interaction chromatography 450

  • see Contents 235–236
  • ammonium sulfate 261, 267
  • application examples 258–266
  • appropriate buffers and salts 241–244
  • buffers 243
  • cleaning in place (CIP) 256
  • column and gel maintenance 256
  • column calibration 246
  • detergents 249
  • ethylene glycol 249
  • gels, syntheses and capacities 239–241
  • hydroxyapatite principles 273
  • immodulised calmodulin 748
  • optimization of running conditions 248–250
  • packing the column 245–246
  • preparation of samples 244
  • recombinant proteins 237, 571
  • salting-out effect 242
  • sample properties 256
  • system set-up 244
  • troubleshooting 257, 266–267

• Hydrophobic interactions 114, 628

• Hydrophobic proteins 807

• Hydrophobic salting-out chromatography 236

• Hydrophobic tail 528

• Hydrophobicity 62, 149, 532, 850

• Hydropore 461

• Hydropore anion-exchange column 459

• Hydroxamic acids 335

• Hydroxyapatite 594, 814

• Hydroxyapatite chromatography

  • see Contents 271–272
  • application examples 292–297
  • back pressures 275, 279, 288, 296
  • capacities 288
  • column packing 280–281
  • commercially-available gels and columns 275
  • cytochrome P-450 287
  • phosphate 279–280
  • removal of detergent 278, 287–288
  • sample preparation 280
  • set-up 279
  • test mixture 284
  • troubleshooting 296–297
  • volatile buffers 280

• 4-Hydroxybenzoic acid 193

• Hydroxylapatite 271

• Hydroxylated polyether copolymer 127

• Hydroxylation 292

• 12′-Hydroxysteroid dehydrogenase 434

• Hygiene 343

• HyperD 315, 787, 791, 807

• Hyper D F 775

• HyperD gels 315, 335, 744, 787, 791–793, 795, 797–798, 807–809

• HyperD media 335

• Hyperglycemia 70

I

• IAM interfacial region 457

• IAMC

  • see Immobilized artificial membrane chromatography
• IDA

  • see Iminodiacetic acid
• EEC

  • see Ion exchange chromatography
• IEF

  • see Isoelectric focusing

• IgA 493

• IgG 482, 484, 487, 493, 496, 823

• IgG₁ 366

• IgG Sepharose 567

• IgM 493

• IMAC

  • see Immobilized metal ion affinity chromatography

• IMAP 373

• IMEC 334

• Imidazole gradient 322

• Imidazole 289, 321, 332, 351, 392, 397

• Iminodiacetic acid 34, 304–324, 334–335, 338, 344, 351, 353, 355, 358–360, 365, 367–370, 372–376, 394, 397, 520, 707, 711, 713, 715, 719

• Iminodiacetic acid 304, 311

• Iminodiacetic acid 707

• Iminodimethyldiacetic acid 335

• Iminodipropionic acid 335

• Immbolized phoshpolipids 457

• Immobilized antibodies 706

• Immobilization of nucleotides

  • see Contents 717
  • connecting a ligand 724–725
  • linking 719
  • new applications for nucleotide gels 726–727
  • purification using nucleotides 719
  • solid phase syntheses using Fractogel EMD Epoxy 726
  • toxicity 720
• Immobilization of

  • 2′, 5′-ADP 759
  • 5′-AMP 759
  • antibodies 718
  • calmodulin 748
  • dehydrofolate reductase 661
  • disulfide 643
  • heparin 747
  • metal ions in IMAC 317–319
  • nucleotide sugars 720
  • phosphate group 720
  • Protein A 718
  • Protein G 718
  • proteins on an azlactone tentacle gel 715
  • support materials 721
• Immobilized artificial membrane

  • chromatography
  • see Contents 449
  • cytochrome P-450 466
  • detergent step gradient 460
  • endcapped 461
  • equipment set-up 464
  • integral membrane protein 464
  • large-scale purification 463
  • preparative-sized column 463
  • protein content measurement 452–453
  • purification examples 453–464
  • sample preparation 451

• Immobilized iminodiacetic acid tentacle gel 715

• Immobilized lipid 464

• Immobilized metal ion affinity chromatography

  • see Contents 301–302
  • antibodies 364
  • application examples 348–371
  • automation 345–346
  • bleeding or leakage 312, 318, 323, 337–338, 342, 364
  • buffer solutions for a pH gradient 329
  • commercially available gels 314–317
  • cytochrome P-450 isozymes 352–354
  • cytochromes c 302, 304–306, 309, 319, 324–325, 327
  • detergents 337, 364
  • eluting agents 321–323
  • enzyme activity, binding, inhibition, stability and topography 308–309
  • Fractogel EMD 316
  • glycoproteins 308
  • HiTrap columns 313
  • IDA 304, 311, 360
  • immunoglobulins 306, 308, 364
  • large-scale preparation of factor IX concentrate 369–371
  • membrane proteins 306, 541
  • metal ions 317–319
  • metalloproteins 306, 318, 372
  • monoclonal antibodies 370
  • new and unusual gels 335
  • NTA 304, 311, 359–360
  • optimizing 334–341
  • photosystems I and II 306, 348
  • pre-bleeding 323, 338, 353
  • principles 303–350
  • recombinant and fusion proteins 307–308, 324, 335, 337, 357–364, 564
  • scale-up 342
  • scaling down 342
  • SMART system 325
  • TED 304, 311
  • tentacles 313
  • test run using a syringe 330–333
  • test run with an FPLC system 327–330
  • test run with an HPLC system 325–327
  • trace enrichment 346
  • troubleshooting 374, 376

• Immune mediator release 633

• Immunoaffinity chromatography 678

• Immunobac 544

• Immunoglobulin G 633

• Immunoglobulins 306, 308, 364

  • see also Antibodies

• Immunomagnetic bead depletion 669

• Immunomagnetically labeled cells 691

• Immunoprotein affinity gels 364

• IMPACT 560, 575

• Inclusion bodies 579, 766

• Incubation time 683

• INdEX 779

• Induction 591–593

• Industrial downstream processes 718

• Industrial preparative separation 875

• Influenza virus 535, 538

• Initial breakthrough concentration 685

• Initial capture 272

• Injection valve 93

• Inlet and outlet ball 94

• Inner mitochondrial membrane 461

• Inorganic matrix 272

• Insect transferrin 363

• Insulin B-chain 193

• Insulin cleaving proteinase 264

• Integral membrane protein 658, 841

  • see Contents 527
  • application examples 537–547
  • bioaffinity chromatography 534, 537
  • biological activity 534
  • cloud point 532
  • critical micellar concentration (CMC) 529–530
  • cytochrome P-450 536, 538
  • detergents for membrane proteins 464, 529–530
  • ELISA 529
  • hydrophobicity 532
  • ion exchange HPLC 534, 536
  • micellar molecular weight 530, 532
  • monoclonal antibodies 544
  • phospholipid 528–529
  • price(s) 533
  • reversed-phase HPLC 534, 538
  • size exclusion HPLC 534, 535
  • troubleshooting 548
  • UV-transparency 533

• Integration 874

• α₁β₁-Integrin 535

• Interfacial enzymes 453

• Interferon 306, 308, 432, 747

• Intermolecular dioxygenases 294, 408

• Intestine 441, 463

• Intestinal peptide transporter 536

• Intramolecular disulfide bonds 641

• Ion exchange buffers 33

• Ion exchange chromatography 594, 598, 624–625

  • see Contents 3–4
  • see also Anion exchange chromatography
  • see also Cation exchange chromatography
  • β-lactoglobulins 51
  • additives 37, 39
  • anion exchangers 18
  • antibodies 76
  • application examples 70–78
  • buffers 34
  • capacity 31–32
  • cation exchangers 21
  • chromatography on zirconia 78–79
  • cytochrome P-450 41
  • detection 48
  • detergent removal 68
  • detergents 38, 63, 67
  • eluting agents 35–37
  • equipment 10–15
  • Fab fragments 76
  • gel itself 16–24, 54–55
  • glycoproteins 69–70
  • HPLC 80
  • hydrophilic–lipophilic balance 64
  • hydrophobicity 62
  • isozymes 41, 72
  • liver microsomes 41
  • optimization 49–52
  • packing the column 46–47
  • particle size and porosity 24–28
  • porosity 6
  • purification of immunoglobulin G₁ 74–76
  • sample preparation 42–44
  • scaling up and down 56–57
  • SNAP 33, 56, 74, 80
  • strong ion exchanger 80
  • system set-up 41–42
  • tentacle ion exchangers 7, 55
  • troubleshooting 77–78
  • UV/VIS-monitors 13
  • weak ion exchanger 81
  • xenobiotics 42

• Ion exchange 273

• Ion exchange HPLC 540

• Ion exchanger 387, 596

• Ion pair reagent 149

• Ionic capacity 80

• Ionic effects 114

• Ionic interactions 114

• Ionic strength 280, 387, 395

• Ionotope 7, 80

• Ion-pair model 153

• IPTG 569, 573, 575, 597, 604

• Irreversible binding 150

• Isocitrate 404

• Isocitrate dehydrogenase 719

• Isocratic formation 387

• Isocratic pump 94

• Isoelectric focusing 70, 73–74, 76, 207, 212, 222, 386–387, 489, 492, 495, 499, 675

• Isoelectric point 114–115, 119–120, 181, 249, 257, 386, 479, 490, 499, 565, 630, 742, 752, 780, 792, 841, 847, 849

  • Immobilized metal ion affinity chromatography 305, 310, 319, 361, 386–389, 392, 396, 400, 404, 406–410, 412
  • Ion exchange chromatography 7, 18–19, 22, 32, 47, 71, 73, 78, 80

• Isoelectric variants 625

• Isoenzymes 272, 292, 404, 462

• Isoforms 292, 404, 624

• Isoforms of monoclonal antibodies 71

• Isopropanol 116, 133

• Isothiocyanate 626

• Isourea 634

• Isourea bond 708

• Isozymes 41, 72

J

• Journal of Chromatography 151

K

• 2-Keto-3-oxy-6-phosphoglucinate aldolase 435

• Kinases 718, 752

• Kininogen 306

• Klenow fragment 609, 611–613

• Kyte–Doolittle plot 62

L

• Laboratory integrated management system 865

• Lactate dehydrogenase 430, 434, 492, 499, 719, 753

• Lactoferrin 306, 318, 431

• β-Lactoglobulins 51

• Lactose-Sepharose 624

• LALLS 94, 137, 238

• LALLS photometer 137

• Lambda repressor 588

• Large proteins 218

• Large scale 57, 72, 156, 306, 360, 369–370, 417, 463, 559–560, 565, 657, 734, 766–767, 769–772, 774, 781, 782, 788–789, 798, 813, 815

  • see also Scaling up

• Large zone 138

• Large zone size exclusion 137

• Large-scale preparation 369

• Large-scale purification 463

• β₁-Latency-associated peptide (β₁-LAP) 435

• LC-GC-online 151

• LDH 489

• Leaching 312

• Leakage 312, 323, 743

• LEC 303

• Lectin activity 629

• Lectin blotting 622

• Lectin(s)

  • see also Agglutinin(s)
  • see also Glycoconjugate(s)
  • see also Cell affinity chromatography
  • agarose gel 680, 687
  • animal 627
  • calcium binding 307
  • carbohydrate combination 678
  • coupling reactions 706
  • galactoside-binding 624
  • general 620–622
  • heparin-binding 623
  • horse gram 317
  • IMAC
  • mistletoe 627
  • plant 627
  • serial lectin affinity 620, 630
  • soybean 684, 688
  • tissue 630
  • Urtica dioica 623
  • wheat germ 628, 673

• Lectin-bound cells 678

• Lectin-containing gels 622

• Lectins and glycoconjugates

  • see Contents 619
  • agglutinins 620
  • application examples 632–634
  • combination with other chromatographies 630
  • glycohistochemical monitoring 632
  • lectin purification modification 626–628
  • lymphocyte migration 632
  • NMR spectroscopy 620
  • standard purification scheme 623, 625
  • sugar receptor purification 632
  • troubleshooting 634–635

• LEF-1 575

• Leishmania membrane protein 536

• Lens culinaris agglutinin 629

• Lentil lectin 401

• Leukaemia cells 355

• Leukocytes 679

• Leupeptin 625–626, 856

• Lewis acid 79, 304

• Lewis base 79, 304, 312, 347

• LHRH 308, 358

• LiChrospher-Diol 472

• LiChrospher Diol 4000 478, 483, 489

• Ligand blotting 626

• Ligand densities 237, 709

• Ligand exchange chromatography 303

• Ligand immobilization 672

• Ligand(s)

  • cell affinity chromatography 668–669, 671–672, 674–676, 678–679, 683–684, 686–689, 692–695
  • chromatography using Strep-tag affinity peptides 823, 827, 831
  • coupling reactions 706–716
  • covalent chromatography 640, 643–646, 648–650, 652–653, 659–660
  • DNA-binding proteins 594, 600–601
  • dye-ligand affinity chromatography 416–445
  • hydrophobic interaction chromatography 236–241, 248, 250–252, 266
  • immobilization of nucleotides 717–720, 719–726
  • immobilized artificial membrane chromatography 453–454, 466
  • immobilized metal ion affinity chromatography 303–306, 311–312, 317–318, 321–323, 349–350, 353, 364–365, 376
  • integral membrane proteins 536, 541–548
  • ion exchange chromatography 8–9, 22, 41
  • lectins and glycoconjugates 620–623, 625–630, 632, 634–635
  • liquid–liquid partition chromatography 471, 486, 490, 494, 498–500
  • miscellaneous biospecific affinity ligands 732–734, 742–754, 757–761
  • scale-up of downstream processing 771–772, 781

• Light chain 366

• Light-harvesting complex proteins 816

• LIMS 865

• Linear flow rate 80, 776

• Linear gradient 809

• Linear velocity 799

• Linking 719

• LiParGel 650 472, 478, 483, 489

• Lipid environment 287

• Lipids 40, 140, 323, 528, 811

• Lipoproteins 733

• Lipoprotein Upase 734

• Liquid–liquid partition chromatography

  • see Contents 469–470
  • antibody difference detection by LLPC 484, 498
  • application examples 488–508
  • chromatograms in LLPC 483
  • column dimensions and scaling up and
  • down 486
  • gels 472
  • IgG 482, 484, 493, 496
  • LiChrospher Diol 4000 478, 483, 489
  • monoclonal IgG 495–498
  • optimizing of running conditions 482
  • partition properties for LLPC 487
  • PEG 8000 480
  • polyclonal IgG 491, 494
  • sample preparation 479
  • surface properties 498
  • surface property comparison by LLPC 497–498
  • system set-up 475
  • troubleshooting 501–502

• Lithium chloride 852

• Lithium salts 851

• Lithiumdodecylsulfate 135

• Liver microsomes 41, 293, 462

• Liver 443

• LIVM values 62

• LLPC

  • see Liquid–liquid partition chromatography

• Loading capacities 809

• Low-angle laser light-scattering 136

• Low angle laser light scattering photometry 238

• LPLC (low pressure liquid chromatography)

  • immobilized metal ion affinity chromatography 324, 330–333
  • ion exchange chromatography 10–11, 13–14

• Lubrol 852–853

• Lubrol PX 43, 530

• Lymphocytes 356, 533, 672, 675, 679

• Lymphocyte migration 632

• Lymphokynes 694

• Lyophilization 217, 846

• Lysozyme 244, 247, 249, 252, 285, 306, 309

• l-Cysteine agarose 645

M

• α₂-Macroglobulin 306, 319

• Macromolecules 289

• Macroporous gels 709

• Macro-Prep 775, 787, 791, 807

• Macro-Prep gels 791–798

• Magnetic activated cell sorter 691

• Magnetically stabilized fluidized bed 692

• Magnetized steel wool columns 691

• Malate dehydrogenase 492, 753

• MALDI-MS 155, 513

• Maltose binding protein 562, 565

• β-D-Maltoside 288

• Mammalian cells 767

• Mammalian galectin-1 633

• Manduca sexta 363

• Mannan 635

• Mannan-binding protein 620, 622

• Mannitol 115

• Mannitol-specific enzyme II of the E. coli 535

• Mannose 634

• Mannose-6-phosphate receptor 634

• Manufacturers 887–894

• Marker proteins 400

• Mass recovery 241

• Mass spectrometer 154

• Mass spectrometry 143, 195, 620, 620

• Mass spectroscopy 140, 142

• Mast cells 676

• Matrix 336, 417, 473, 709

• Maximizing resolution 288

• MDH 489

• Measles virus 536–537

• Mecuric reductase 431

• MedLine 151

• Membrane affinity chromatography surfac 454

• Membrane antigen (Plasmodium falciparum) 536

• Membrane filters 285

• Membrane glycoprotein antigen 535

• Membrane multiprotein complex 814

• Membrane protein complexes 835

• Membrane protein solubilization 854

• Membrane protein 143, 222, 273, 306, 410, 453, 536–538, 806–807, 811

• Membrane 450

• Membranes (E. coli) 536

• Mercaptoethanol 854

• 2-Mercaptoethanol

  • covalent chromatography 641–642, 648–650, 661–663
  • dye-ligand affinity chromatography 430–431, 433–434, 443–444
  • hydrophobic interaction chromatography 243
  • immobilized artificial membrane chromatography 456, 466
  • immobilized metal ion affinity chromatography 303, 320, 321, 334, 376
  • ion exchange chromatography 40, 77
  • lectins and glycoconjugates 621, 623, 626
  • miscellaneous biospecific affinity ligands 739, 841, 854
  • recombinant proteins 576
  • size exclusion chromatography 98–99, 105

• Merthiolate^® 627, 737

• Metabolic pathways 841

• Metal ion 486

• Metal ion leakage

  • see Bleeding

• Metal ion transfer 372

• Metalloproteases 855

• Metalloproteins 306, 318, 372, 827

• Metastasis formation 632

• Methanol 132, 291

• Methylation 614

• Methylation footprint 612

• Methyl-Sepharose 748

• MHC class II-peptide antigen complexes 535

• Micellar molecular weight 530, 532

• Micelle size 116

• Microbial contamination 402, 812

• Microbial growth 59, 310

• Microbiological growth 194

• Microbore columns 154

• Micrococcus luteus 532

• Microfiltration 767

• Microheterogeneity 142

• Microsomal membrane proteins (rat liver) 536

• Microsomal proteins 462

• Microsomes 42

• Microviscosity 457

• MiniMACS 695

• Mistletoe lectin 627

• MIT 372

• Mixed disulfide bonds 641

• Mixed-bed gels 143

• Mobile phase system 180

• Mobile phase 5, 465

  • see also Buffer(s)

• Mode of branching 630

• Model membranes 140

• Modifier 114, 457

• Modifying agents 320

• Moduline acrylic 779

• Moduline ss 779

• Molecular maquettes 138

• Molecular mass determinations 93

• Molecular size 90

• Molecular weights 90

• Moloney murine leukemia virus 538

• Mono and dinucleotides 307

• Mono P

  • chromatofocusing 390–391, 394, 397–398, 400, 402, 406–407, 409–410, 412
  • DNA-binding proteins 599
• Mono Q

  • buffers and additives 848–849
  • DNA-binding proteins 599
  • hydroxyapatite chromatography 297
  • immobilized metal ion affinity chromatography 320, 355
  • integral membrane proteins 542
  • ion exchange chromatography 15, 19, 26, 29–30, 36, 40, 47, 49–50, 53, 57–59, 66–68, 72–74
  • recombinant proteins 577–578
• Mono S

  • displacement chromatography 519
  • DNA-binding proteins 589, 597, 599, 604–606
  • ion exchange chromatography 15, 21, 26, 30, 40, 47, 58–59, 67, 71–72

• Monoamine transporter 535

• Monoclonal antibodies 370, 544, 672, 682, 823

• Monoclonal IgG 495, 498

• Monoclonal IgG2b 496

• Monocytes 672

• Monohydrogenphosphate 279

• Mononucleotides 307

• Monooxygenases 292

• Motional properties 457

• MPLC (medium pressure liquid chromatography)

  • see FPLC (fast protein liquid chromatography)
• MS

  • see Mass spectrometry

• Mucin 621, 627, 631–632, 634

• Multicolumn methods 866

• Multimedia-HPLC 875

• Multi-receptor attachment 684

• Multi-tasking systems 865

• Murine IgG₁ 365

• Muscarinic acetylcholine receptor 535

• Mycoplasma gallisepticum 535–536

• Mycoplasma hyopneumoniae 538

• Myc-tag 826

• Myelin glycoprotein P₀ 538

• Myeloma IgG 497

• Myoglobin 78, 244, 247, 252, 306, 309, 489, 790

• Myosin 432

• Myosin light chain kinase 747

N

• N-(2-pyridyl-methyl)glycine 335

• N-(4-vinylbenzyl)-imino]diacetic acid 335

• N ⁵-methyltetrahydromethanopterin: coenzyme M 535–536

• N-acetyl-cysteine 571

• N-acetylgalactosamine 620, 629, 681

• N-acetylneuraminic acid 632

• N-acetylphosphatidylethanolamine synthase 453, 455, 466

• N-hydroxysuccinimide 622, 640, 659, 719, 744, 773

• n-octyl-β-D-glucopyranoside 116

• NAD 601, 757

• NAD⁺ 429, 434, 747, 753–754, 718–719

• NADH 294, 430, 435, 440, 450, 499, 535, 718, 733, 747, 752–754, 757–758

• NADH oxidase 430, 535

• NADH⁺ 499

• NADH/NAD⁺ 718

• NADP 6, 67, 757

• NADP⁺ 435, 660–661, 718–719, 753,

• NADPH 431, 718, 733, 747, 752–754, 756–759, 851

• NADPH cytochrome P-450 reductase 754

• NaDPH/NADP⁺ 718

• Nanofiltration 767

• NAPES 466

• Naphthalene dyes 428

• Natural killer cells 678, 688

• Negative purification 319

• Negative selection 679, 688

• Negative selection panning 694

• Neopentyl 240

• Neuraminidase 679–680

• Neuronal cells 676–677

• Neuropeptides 195

• NHS

  • see N-hydroxysuccinimide

• Ni²⁺-IDA 360

• Ni²⁺-NTA 360

• Ni-NTA 316

• Ni-NTA agarose 566

• Ni-NTA Silica 316, 566

• Ni-NTA superflow 566

• Nitric acid 129

• Nitroloacetic acid 304, 306–308, 311–313, 316–317, 321, 334, 344, 358–363, 373–374, 592

• NMR 142, 471, 573, 588, 814

• NMR spectroscopy 620

• Non-denaturing 852

• Non-fusion recombinant proteins 360

• Nonidet P40 530

• Non-ionic detergents 273, 404

• Non-polymeric focusing 395

• Non-selective desorption 457

• Non-specific binding 672, 685

• Non-specific cell adsorption 670

• Non-specific cell entrapment 692

• Non-specific ionic interactions 319

• Novarose 364

• NP-40 410, 529

• NTA

  • see Nitroloacetic acid
• Nuclear magnetic resonance

  • see NMR

• Nucleic acid binding proteins 733, 738

• Nucleobase-linked nucleoside phosphates 721

• Nucleosid diphosphate kinase 434

• Nylon wool column 695

O

• Oat phytochrome A 827–828

• Object linked embedding 865

• Octyl glucoside 320, 853–854

• Octyl-glucopyranosid 834

• Octylglucoside 529–531

• OLE 865, 875

• Olfactory neuronal transcription factor 614

• Oligomer 123

• Oligomerizations 120

• Oligosaccharides 621

• OmpA signal sequence 571

• OMPs (E. coli) 536

• OMPs of Haemophilus influenzae 535

• On-column cleavage 576

• Oncoprotein bcl-2 537

• One-hybrid system 588

• On-line dynamic light scattering detector 94

• On-line light scattering detector 93, 143

• Online low-angle laser light scattering detector 94

• On-line viscometer 93

• Operating system 865

• Operator 588

• O-phosphoserine 335

• Optical filter 328

• Optical flow-through cell 93

• Optimization 188, 810

• Organic bonded phases 125

• Organic compounds 291

• Organic modifier 546

• Organic polymers 92

• Organic solvent elution 453

• Organic solvents 40, 127, 131, 280, 403, 457

• Organosilane 621

• Orotate phosphoribosyltransferase 432

• OS/2 866

• Osmotic shock 561

• Outer membrane proteinase 264

• OVA 489

• Ovalbumin 50, 252, 635, 798, 800

• Ovalbumine 805

• Ovomucoid 621, 628

• Oxamate 499

• Oxirane 708, 724

• Oxirane-activated matrices 241

P

• P-450

  • see Cytochrome(s) P-450
• Packing of column(s)

  • see Column(s), packing of

• Packing reservoir 281, 399

• Packing state 131

• Packings 240

• Pancreatic PLA₂ 454, 466

• Panning 669, 695

• Papain 650, 855

• Papain-digestion 77

• Particle size 178, 281, 809

• Partition coefficient 471

• Pass through 282

• Pass through peak 465

• PBE

  • see Polybuffer exchanger

• PC data use 865

• PD10 column 117

• Peak broadening

  • chromatography on porous matrices 787, 789, 809
  • immobilized metal ion affinity chromatography 338, 346
  • ion exchange chromatography 26, 28, 46, 49, 80
  • reversed-phase chromatography 188, 202
  • size exclusion chromatography 118, 125, 128, 130, 132, 189

• Peak capacity 92

• Peak labeling 874

• Peak splitting 131

• Peak volume 112

• PEEK 154, 200, 790–791, 801, 803, 824, 864

• Pefabloc SC 855–856

• Pefabloc^® 739

• PEG 8000 480

• PEG

  • see Polyethylene glycol

• Penicillium amagasakiense 53, 72

• PEPcK

  • see Phosphoenolpyruvate carboxykinase

• PEP-dependent phosphotransferase system> 535

• Pepsin 392

• Pepstatin 856–857

• Peptide hydrolase 653

• Peptide mapping 208

• Peptide maps 202

• Peptides 121, 138

• Peracetic acid 693

• Perchlorate 183

• Perforins 354

• Perfusion chromatography 80, 176, 787–788

  • see also Chromatography on porous matrices and POROS gels

• Peripheral basophils 676

• Peripheral blood cells 677

• Peripheral blood lymphocyte 695

• Peripheral membrane protein 293, 454, 528

• Periplasmic proteins 561

• Peristaltic equipment 789

• Peristaltic pumps 670

• Peroxidase 489

• pH

  • adjustment of for size exclusion chromatography runs 114, 849
  • analysis of for protein folding by size exclusion 141–142
  • changing pH for optimization of hydrophobic interaction chromatography 249, 850
  • changing pH for optimization of membrane protein separation 810
  • choice of for 5 ′–AMP and 2′, 5′-AMP runs 757
  • choice of for affinity chromatography 850–851
  • choice of for covalent chromatography 651
  • choice of for immobilized calmodulin 750
  • descending pH gradients in immobilized metal ion affinity chromatography 327, 329, 338
  • for carrier support of pH in displacement chromatography 513, 515
  • for influence of on binding to Mono Q 848
  • for protein folding and stability with Strep-tag affinity 826
  • for use in buffers 842–843, 845–847
  • pH gradients in ion exchange chromatography 52, 66, 71, 73, 76
  • pH limits in ion exchange chromatography 32, 36
  • pH of aqueous components for reversed-phase chromatography 180
  • pH regions in chromatofocusing 356–357, 849
  • silica-based sorbent choices for reversed-phase chromatography 186

• pH-changes 290

• pH interval 408

• pH monitor 327

• Pharmalyte 393–394, 849

• Phase separation 853

• Phases 452

• Phenol red 400

• Phenol(resorcinol)-sulfuric acid assay 628

• Phenyl sepharose 570

• Phenylarsine oxide 568

• Phenylglyoxal 622

• Phenylmercury salts 861

• pH-gradients 387

• Phosphatases 306, 752

• Phosphate buffered saline 678

• Phosphate 115, 127, 279–280, 843

• Phosphatidylcholine 451, 453–454

• Phosphatidylethanolamine 451, 453–456

• Phosphoacetylmuramylpentapeptide translocase 532

• Phosphoamino acids 322

• 1,6-Phosphoaminohexyl spacer 759

• Phosphoenolpyruvate carboxykinase 260–264

• Phosphofructokinase 416

• 6-Phosphoglucanate dehydrogenase 754

• 6-Phosphogluconate dehydratase 433

• Phosphoglycerate kinase 434

• Phospholipase A₂ 454

• Phospholipid 528–529

• Phospholipid bilayer 451

• Phosphopeptides 154, 318

• Phosphoproteins 306, 322

• Phosphoramidon 856

• Phosphoric acid 843

• Phosphorlyated nucleotides 752

• Phosphorylation 634

• Photo diode array detector 93

• Photoautotrophic organisms 294

• Photosynthic membrane proteins 806–809

• Photosystem 67–68, 121, 123–124, 136–137, 296, 306, 535, 7, 828

• PS 1291, 295, 297, 348–350, 814, 816–818, 824

• PS II 294–295, 348–350

• Photosystem II 294

• Phthalocyanine dyes 428

• Physiological conditions 842

• Phytochrome 431

• pI

  • see Isoelectric point

• Pigment 121

• Pilot chromatographic studies 464

• Pinpoint peptide 563, 568

• PIPES 291

• Piston seal rinse 94

• pK _a 33, 40, 842

• pK _a-values 52, 66

• PLA₂ 456, 458–459, 461, 466

• Placenta 623, 626, 631–633

• Plasma 370

• Plasma membrane proteins 543

• Plasma membrane proteins (human) 537

• Plasma membranes (liver)/morris hepatoma 535–537

• Plasma protein 511

• Plasmacytoma cells 677

• Plasminogen 434

• Plasminogen activator 306

• Plasmodium vivax 658

• Platelets 677

• Platelet-derived growth factor receptor 535–536, 538

• Plugging 811

• Pluronic F68 363

• PMSF 37, 39, 41, 43, 293, 320, 348, 359, 361, 754, 841, 855–856

• Polarity of solution additives 859

• Polishing (Final step) 150, 272, 287, 323, 341, 348, 653, 742

• Poly(2-hydroxyethyl methacrylate 694

• Polyacrylamide 621, 672–673, 706

• Polyamine graft copolymer 694

• Polybuffer 393–394, 405, 849

• Polybuffer exchanger 94, 118, 258–259, 390–393, 397, 399–403, 406, 408, 410, 411, 861

• Polychlorosilanes 174

• Polyclonal IgG 491, 494

• Polyclonal immunoglobulins 365

• Polyethylene 471

• Polyethylene glycol 39, 677, 860

  • dye-ligand affinity chromatography 437–439, 442–444
  • immobilized metal ion affinity chromatography 334, 336, 373, 376
  • liquid–liquid partition chromatography 471, 474, 476–480, 483–487, 489–497, 499, 501

• Polyethylene glycol precipitation 336

• Polyethyleneimine 594

• Polyethylenimine-modified silica gels 389

• Polyglycidyl methacrylate chains 710

• Polyglycidylmethacrylate grafted-tentacle gel matrix 715

• Polyhydroxymethacrylate 106

• Polymethacrylate 706

• Polymyxin B sulfate 561

• Polynucleotide phosphorylase 432

• Polyols 38

• Polyoma virus medium size tumor antigen 537

• Polyoxyethylene alkyl ether 530

• Polypeptide 141

• Polysaccharides 620

• PolySep 106

• Polystyrene 673

• Polystyrol 706

• Polyvinyl pyrrolidone 623

• Porath 310

• Porcine growth hormone 185

• Pore size

  • buffers and additives 860
  • cell affinity chromatography 669, 680
  • chromatofocusing 398
  • chromatography on porous matrices 787–788, 791
  • coupling reactions 709
  • covalent chromatography 644
  • displacement chromatography 518
  • dye-ligand affinity chromatography 417
  • hydroxyapatite chromatography 280, 285
  • immobilized metal ion affinity chromatography 313–316
  • integral membrane proteins 546
  • ion exchange chromatography 8, 27, 77
  • liquid-liquid partition chromatography 472–473, 486
  • non-porous media 786
  • recombinant proteins 577
  • reversed-phase chromatography 156, 158, 160, 168, 172, 175, 177–179, 184–186, 203–204, 206, 210–212, 214–216, 218, 220, 222
  • scale-up of downstream processing 775, 781
  • size exclusion chromatography 94–95, 112–113, 117, 128, 143

• Pore size distribution 143, 791

• POROS 50, 775, 787, 791, 807

• POROS gels 787, 789–809

• POROS HS 820

• POROS ion exchangers 55

• POROS R2 819

• POROS S 820

• Porosity 6, 16, 24, 26, 32, 79, 92, 128, 176, 313, 646, 714, 735, 749, 755, 758

• Porous glass 113

• Porous matrices

  • see Contents 785–786
  • advantages and disadvantages 788
  • antibody purification and screening 823
  • application examples 813–823
  • back pressure 788, 798, 806, 811
  • binding capacity 788
  • column dimensions 808
  • column packing 800
  • equipment 789
  • flow rates 788–790
  • gels on porus matrices 791, 798
  • general utility 788
  • optimization 810
  • particle sizes 809
  • pressure 799
  • sample preparation 800
  • scaling up and down 808
  • storage 812
  • troubleshooting 824
  • work stations 810

• Positive selection 678–679, 694

• Post-run analysis 866

• Post-translational modifications 559

• Potassium chloride 405

• Pre-bleeding 323, 338, 353

• Pre-charging 323

• Precipitation effect 850

• Pre-column 154

• Pre-packed column 280

• Preparative columns 290

• Preparative-sized column 463

• Preparing a buffer 846

• PreScission protease 563

• Presence 854

• Preservatives 59

• Pressure 799

• Pressure drop 687, 692

• Pressure variations 94

• Price(s)

  • see Cost(s)

• Primary amines 321, 851

• Principles 706

• Probond 566, 592

• Processing capacity 685, 690–691

• Procion Brown MX-5BR 428

• Procion Green H-4G 428, 436

• Procion Navy H-ER 436

• Procion Red 486

• Procion Red H-E3B 428

• Procion Red MX-2B 436

• Procion Yellow MX-8G 436

• Procion Yellow MX-GR 436

• Procion^® dyes 428

• Procollagenase 435

• Proinsulin 308, 358

• Prokaryotes 767

• Promoter 588

• Promoter operator 591–593

• Properdin 734

• PROSEP-Heparin 744

• Prostaglandin H₂ synthase 535

• ProSys 777

• Protease inhibitors 41, 291, 306, 337, 623, 750, 827, 841, 854, 856

• Proteasome 558

• Protecting 841

• Protein A 365, 563, 672–673, 678, 706, 718, 772, 798

• Protein A affinity resins 772

• Protein A-Sepharose 633

• Protein binding 806

• Protein binding capacity 709

• Protein bioactivity 826

• Protein degradation 404

• Protein design 138

• Protein determination 247, 262

• Protein folding 826

• Protein G 365, 718, 798

• Protein KW 101

• Protein PAK 60 100

• Protein splicing 560

• Protein structure 826

• Protein synthesis factors 733

• Protein test mixture 285

• Protein tyrosine phosphatase 536

• Protein-DNA interactions 587

• Proteoglycans 354, 629, 742

• Proteoglycan core proteins 622

• Proteoliposomes 140

• Proteolytic degradation 582–583

• Proteolytic enzymes 131

• PS I- and II-monomers 297

• PS I-trimer 297

• Pseudo-bioaffinity ligands 707

• Pseudomonas aeruginosa 264

• Pulsation 94

• Pulse damper 129

• Pulse detergent gradient 453, 455

• Pulse gradient 454

• Purification by HIC 264

• Purification tag 590

• Purity 846

• Pyrogen testing 693

• Pyroglutamyl aminopeptidase 653–654

• Pyrrolidonecarboxylyl peptidase 653

Q

• Q Sepharose HiLoad 814

R

• Ragi bifunctional inhibitor 571

• Rat lens 535

• Rat liver microsomes 41

• Reaction center from Rhodopseudomonas spheroides 535

• Reaction center Photosystem I and II 535

• Reaction intermediates 816

• Reactive dyes 426–427

• Reciprocating pistons 94

• Recombinant

  • chromatofocusing 407–410
  • chromatography on porous matrices 813, 815–816
  • chromatography using Strep-tag affinity peptides 826–827, 830–833
  • DNA–binding proteins 588, 597
  • hydrophobic interaction chromatography 235, 238
  • immobilization of nucleotides 719, 726
  • immobilized metal ion affinity chromatography 306–308, 324, 335, 337, 357, 360, 363–364, 368–369, 377
  • miscellaneous biospecific affinity ligands 739, 747, 754, 755, 759
  • recombinant proteins 557–586
  • reversed-phase chromatography 208–209, 215–217, 228–229
  • scale-up of downstream processing 766, 773
  • size exclusion chromatography 137–138, 150

• Recombinant peptides and proteins 215

• Recombinant protein production 826

• Recombinant proteins

  • see Contents 557
  • application examples 569–579
  • contaminating proteins co-eluting with the fusion protein 582
  • disulfide bonds 571
  • expression systems 559
  • failure to bind to affinity resins 583
  • formation of inclusion bodies 579
  • fusion partners 559–560, 562–563, 568
  • fusion proteins 306, 308
  • gel matrices 564, 566–567
  • his-tagged proteins 564
  • on-column cleavage 576
  • sample preparation 561
  • troubleshooting 579
  • usefulness of HIC 237

• Recombinant Tick anticoagulant peptide 813

• Recovery 275

• Red blood cells 463

• Redox titrations 140

• Reductase 754

• Reductive amination 626–627

• Refraction detector 513

• Regeneration protocol 345

• Regeneration

  • see Gel(s), regeneration

• Regulatory support file 706

• Remazol^® dyes 428

• Renaturation 560

• Renex 287

• Replenine 371

• Repressor gene 591–593

• Repressor purification 589

• Reproducibility 454, 457

• Reservoir 11

• Resin(s)

  • see Gel(s)

• Resolution capability 409

• Resolution 80, 275, 404, 788, 808

• Respiratory syncytial virus 537

• Restriction endonuclease 308, 360

• Retinol-binding protein 308

• Reverse transcriptase 306

• Reversed IMAC 372

• Reversed-phase 517

• Reversed-phase chromatography

  • see Contents 147–148
  • alkyl chain length 177–178
  • angiotensin II 193
  • application examples 195–223
  • carbohydrate microheterogeneities 217
  • column conditioning 187
  • column lifetime 191
  • column-ovens 155
  • columns 157
  • desalting 217
  • desalting in reversed-phase 217
  • detectors for reversed-phase 154–155
  • endcapping 174
  • flow rates 183
  • glycoproteins 220
  • hydrophobicity 149
  • large proteins 218
  • MALDI-MS 155
  • membrane proteins 222
  • optimization 188
  • particle size 178
  • pore size 178
  • preparation of eluents 185–187
  • principle 149–153
  • recombinant peptides and proteins 215, 217
  • removal of detergents 217, 223
  • reversed-phase column cleaning 190
  • seoaration of prtoeins 450
  • silicas 174
  • stationary phase 156–180
  • trace enrichment 217
  • troubleshooting 194, 196

• Reversed-phase HPLC 545

• Rhinovirus receptor 537

• Rhodopsin (large hydrophobic peptides) 538

• Ribonuclease A 252, 285–286

• Ribonuclease 306, 401

• Ribosomal proteins 202

• Ribosyl moiety 720

• Ricinus communis 620

• Ricinus communis agglutinin 629, 633–634

• Rigid hydrophilic polymer 109

• RNA polymerase 579, 588, 608, 612–614, 739, 741

• RNA polymerase holoenzyme 608

• RNaseA 826

• Rosetting 669, 679, 695

• RPC

  • see Reversed-phase chromatography

S

• S Sepharose 605

• Saccharomyces cerevisiae 305, 739

• Salts 291

• Salt gradients 454

• Salting out 114

• Salting-in effect 850

• Salting-out effect 850

• Sample application 411

• Sample loading limits 188

• Sample loop 13, 94, 282

• Sample preparation, for

  • chromatofocusing 398
  • chromatography on porous matrices 800
  • covalent chromatography 737, 739, 741–742, 750, 755, 759
  • dye-ligand affinity chromatography 437
  • hydroxyapatite chromatography 275, 280
  • immobilized artificial membrane chromatography 451
  • immobilized metal ion affinity chomatogr. 319, 325–326, 330, 332, 336
  • ion exchange chromatography 41
  • liquid-liquid partition chromatography 479
  • recombinant proteins 561, 641, 646, 652–653
  • reversed-phase chromatography 148, 155, 187
  • size exclusion chromatography 116

• Sample rotator 621

• Sample viscosity 741

• Sandwich formation 634

• Saralasin 193

• Sawtooth gradient 403, 811

• Scale-up of downstream processing

  • see Contents 765
  • buffers for large scale application 774
  • column bed volumes 780
  • column dimensions 780
  • for large-scale applications 772, 774–775
  • microfiltration 767
  • price(s) 774
  • scale-up 765, 776
  • selection of chromatographical methods 771

• Scale-up of downstream processing 765

• Scaling down 57, 342, 486–487, 690, 766, 808

• Scaling up 54, 56–57, 124, 254, 310, 342, 364, 406, 486–487, 652, 690, 694, 765–767, 769–770, 773–777, 779, 781–782, 788, 808–809, 813, 820, 822, 831, 833, 872, 875

  • see also Large scale

• Scanning densitometry 459

• Scouting run function 872

• Screening of dye-ligands 442–443

• SDS 127, 131, 288, 852–853

• SDS-PAGE 452, 842

• SEC

  • see Size exclusion chromatography

• Secondary amine 708, 851

• Selectins 622, 845

• Selectivity 511

• Self-aggregation 852

• Self pack device 801

• Self Pack 803

• Self-packing

  • see Column(s), packing of

• Semliki Forest virus 533

• Sendai virus 531, 535–538

• Separation criteria 386

• Separation of monomers and dimers 491

• Sephacryl S 103, 200 656

• Sephacryl 104, 775

• Sephadex G-25, 200, 648, 673

• Sephadex 620, 775

• Sepharose 313, 626, 775

• Sepharose 4B 673

• Sepharose 6B 673

• Sepharose 6MB 672–673

• Sepharose CL-2B 623

• Sepharose Fast Flow 313

• Sepharose High Performance 313

• Sepharose HP 775

• Sequencing 842

• Serine proteases 855

• Serine-dependent proteases 354

• Serum albumin 491

• Serum proteins 306, 733

• Serum 364, 490

• Sex hormone-binding globulin 490

• Shape 120

• SHBG 489

• Shear rate 686

• Sialic acid 631–632

• Sialyltransferase 726

• Silane 859

• Silanol groups 114

• Silica subsurface amines 461

• Silica 117, 174, 314, 517

• Silica-based gels 313

• Site shadowing 685

• Size exclusion 93, 112, 849

• Size exclusion chromatography

  • see Contents 89–90
  • application examples 134–143
  • buffer exchange 95
  • calibration 94
  • choice of buffer 114
  • column and gel maintenance 129–133
  • column equilibration 121
  • delipidation of membrane proteins 140–141
  • desalting 90
  • detergent 94, 120, 123, 136
  • detergent exchange 140
  • elution modes in size exclusion 125, 127
  • equipment for size exclusion 93
  • fast desalting 105
  • gels for gel filtration, gel permeation and size exclusion 91, 112–113, 787
  • LALLS photometer 137
  • molecular weight determination 134–137
  • optimization 122–125
  • packing the column 118
  • sample loading, washing, elution 122
  • sample preparation 116–117
  • storage of size exclusion column 133
  • system set-up 116
  • troubleshooting 134–135
  • UV detector 93, 137
  • void volume 91

• Size exclusion HLPC 537

• Sjögren's syndrome 497

• Slurry 11

• Sluyterman 387

• Small zone 138

• SMART system 117, 325

• sn-1.2-diacylglycerol kinase 535

• Snake PLA₂ 457–458

• SNAP 33, 56, 74, 80

• Socket valves 94

• Sodium acetate 852

• Sodium ascorbate 409, 857

• Sodium azide 133, 292, 693, 861

• Sodium chloride 405, 851

• Sodium cholate 65, 288, 530

• Sodium deoxycholate 530

• Sodium dodecylsulfate 321, 529–530

• Sodium perchlorate 183

• Sodium taurodeoxycholate 530

• Soft release avidin 591

• Software 11, 15, 451, 681, 740, 810

  • see also Computer chromatography software 863–876

• Software packages 868

• Software updates 875

• Software validation protocol 875

• Solenoid valves 15

• Solid phase syntheses 726

• Solubility 386

• Solubilization 529

• Solubilization buffer 465

• Solubilizing 464, 841

• Soluble amphiphiles 852

• Solute 5

• Sonication 831

• SOTA phase 112

• Source 30, 775

• SOURCE 787, 791, 807

• Soybean lectin 684

• Spacer arm 688

• Spacer displacers 515

• Spacer molecules 417

• Spacer technologies 709

• Spacer 336, 707, 723

• Specific resolution 112

• Specific site 606

• Spectral properties 852

• Speed-vac 217

• S-peptide 562

• Sperm–egg recognition 632

• Splenocytes 691

• S-protein 826

• S-protein agarose 566

• Stacking effects 428

• S-tag 826

• Standard run 800

• Standards for HPSEC 119

• Stationary phase(s)

  • see also Gel(s)
  • chromatofocusing 389–390, 392, 402, 406–407
  • covalent chromatography 640
  • displacement chromatography 506, 508–511, 513, 515–517
  • hydrophobic interaction chromatography 257
  • immobilized artificial membrane chromatography 450, 454
  • immobilized metal ion affinity chromatography 304
  • ion exchange chromatography 5–11, 13, 15–17, 22–29, 31–33, 38–41, 43, 45, 47, 52, 54–60, 62, 64–67, 69, 71, 74–80
  • liquid–liquid partition chromatography 471, 475, 478, 481, 484, 488–489, 502
  • reversed-phase chromatography 149–154, 156, 175–177, 188, 199, 202
  • size exclusion chromatography 91, 95, 114, 128

• Steel 790

• Stem cells 676, 678

• Stem cell isolation 695

• Step gradient detergent elution 453

• Step gradient 288, 809

• Steric accessibility 252

• Steric anion exchange chromatography 273

• Sterile environment 692

• Steroid receptors 733

• Stokes radius 118, 141

• Storage 812

• Strep-peptide 562, 565

• StrepTactin 830

• Strep-tag 830, 833

• Strep-tag II 830

• Strep-tag affinity peptides

  • see also Contents 825
  • membrane protein complexes 835
  • metalloproteins 827
  • practical performance 830–833
  • scale-up 833
  • StrepTactin affinity columns 831
  • troubleshooting 834–835

• Streptavidin 562–563, 565–566, 568, 591, 595, 600, 827, 829–830, 833–834

• Streptavidin tag 591

• Streptavidin-agarose 566

• Streptococcus suis 658

• Stromelysin (metalloendopeptidase) 434

• Strong ion exchanger 80

• Submandibular and parotid gland (rat) 536

• Submaxillary mucin 631–632

• Substance P neuropeptide receptor 537

• Substance P receptor 543

• Substrate composition 841

• Subtilisin 309

• Sucrose density gradient 297

• Sucrose synthase 309

• Sucrose 115, 859

• Sugar phosphate transporter (E. coli) 537

• Sugars 720

• Sugar-specific antibodies 620

• Sulfations 627

• Sulfhydryl blocking agents 654

• Sulfhydryl reagents 291

• Sulfhydryl reagents 40

• Sulfobetain 530

• Sulfobetaines 529

• Sulfone bridges 707

• Sulfone-activated sepharose 626

• Sulphonated polystyrenes 118

• Sulphur 237

• Sumner 620

• Supercomplexes 816

• Superdex 200 478, 483

• Superdex Peptide 101

• Superdex 103, 117, 139, 775

• Superflow 316

• Superformance 779, 802

• Superloop 13, 282, 791

• Superose 102, 127, 313

• Superoxide dismutase 306, 734

• Superparamagnetic beads 373

• Suppliers 887–894

• Support 473, 875

• Surface active agents 852

• Surface chemistry 807

• Surface hydrophobicity 237

• Surface properties 498

• Surface topography 309

• Surfactants 852

• Swelling behavior 23

• SynChropak GPC 107

• Synechocystis 814, 295

• Syntheses

  • chromatography on porous matrices 791, 818, 819
  • coupling reactions 706, 708, 710–715
  • covalent chromatography 643, 650, 659
  • DNA-binding proteins 588, 595, 602
  • hydrophobic interaction chromatography 239, 241, 252
  • immobilization of nucleotides 723–724, 726–727
  • immobilized metal ion affinity chromatography 313, 336, 348, 363, 373
  • miscellaneous biospecific affinity ligands 733, 739, 742, 745–747, 752
  • reversed-phase chromatography 156–174

• Syringe 282

• Syringe operation 330

T

• T cells 679, 687–688

• T helper 694

• T7 peptide 563

• T8 antigen 533

• Tailing 507

• Talon 566

• Tandem column 334

• Tandem 323

• Tannins 623

• Taurine 280, 291, 395

• Taurodeoxycholate 529

• TCA precipitate 453

• TCA precipitation 453

• TEA 217

• TED 304, 306, 309, 311–312, 316–317, 334–335, 350–352, 372, 374, 622, 632, 690

• TED-Agarose 316

• Temperature effects 853

• Temperature

  • buffers and additives 845, 847, 850, 852, 853
  • cell affinity chromatography 670, 674, 682, 684, 693, 697
  • chromatofocusing 389, 395–396
  • chromatography on porous matrices 806, 811–812, 814
  • chromatography software 872
  • chromatography using Strep-tag affinity peptides 827
  • coupling reactions 714–715
  • covalent chromatography 643, 646, 658, 663
  • displacement chromatography 516–517
  • DNA-binding proteins 595, 597, 602, 608
  • dye-ligand affinity chromatography 428–429, 436, 438–439
  • hydrophobic interaction chromatography 239, 245–246, 248, 256, 267
  • hydroxyapatite chromatography 275, 279, 284, 289–290, 292
  • immobilized artificial membrane chromatography 451–452
  • immobilized metal ion affinity chromatography 321, 325, 334, 336, 339, 353
  • integral membrane proteins 532
  • ion exchange chromatography 11, 33, 44, 46, 49, 51, 66
  • lectins and glycoconjugates 621
  • liquid-liquid partition chromatography 472, 474–475, 477, 480–483, 488
  • miscellaneous biospecific affinity ligands 734, 743, 748, 750, 758
  • recombinant proteins 560, 576, 582
  • reversed-phase chromatography 155, 178, 183–185, 188–190
  • scale-up of downstream processing 769
  • size exclusion chromatography 93, 118, 122, 125, 130, 132–134, 137, 142

• Template chromatography 742

• Tensides 852

• Tentacle ion exchangers 7, 55

• Tentacles 113, 313

• TEPA 344

• Test mixture 284

• Test separation 400

• Tetanus toxin 309

• Tetrabutylammonium salts 182

• Tetracyclines 307, 372

• Tetraethylenepentamine 344, 364

• Tetrahydrofolate reductase 433

• Tetrahydrofuran 629

• TFA 153, 181–182, 217

• T-helper cells 678–679, 688

• Theoretical plates 122, 128, 808

• Theoretical titration curve 843

• Thesit 530

• THF 304, 306, 309, 311–312, 316–317, 334–335, 350–352, 372, 374, 622, 632, 690

• Thimerosal 652, 751, 861

• Thiobond 567

• Thiobond Resin 593

• Thioether bridges 707

• Thiol (SH-) reagents 854

• Thiol covalent chromatography 640–641

• Thiol functional group 641

• Thiol oxidation 651

• Thiol proteases 855

• Thiophilic adsorption 707

• Thiophilic adsorption chromatography 237

• Thiopropyl Sepharose 648, 659

• Thiopropyl Sepharose 6B 645

• 2-Thiopyridone 242, 648

• Thioredoxin 309, 563, 593

• Thrombin 590

• Thrombin^b 563

• Through-flow pores 787

• Thylakoid membrane 294, 814

• Thymidine kinase 308

• Thymocytes 677

• Thyroglobulin 487

• Tick-borne encephalitis virus 535, 538

• Tissue lectin 630

• Titration 396

• Titration curve 842–843

• TLCK 856

• TLPK 856

• T-lymphocyte 694

• TNB-thiol gel 645

• Topoisomerase II 739

• Toxic 39, 41, 317, 475, 589, 592, 594, 625, 714, 720, 834, 854–855, 859

• Toyopearl 108

• Toyopearl HW 113

• Trace enrichment 217, 339, 346

• Training courses 875

• Transferases 752

• Transferrin 363, 366, 487, 489–490, 499

• Transmembrane pressure 768

• Treatment of glass surfaces with silane 859

• TREN-Novarose gel 365

• Trichlorbutanol 861

• Trichloroacetic 452

• Triethylamine 621

• Triethylammonium acetate 183, 186

• Trifluoracetic acid 132, 153

• Triglyceride lipase 734

• Trimethylamine 852

• Trimethylchlorosilane 174

• Tri-n-butyl phosphate 370

• Tris 392

• Tris (hydroxymethyl) amine 672

• Tris(2-aminoethyl)amine 335, 364

• Tris(carboxymethyl)ethylenediamine

  • see TED

• Trisacryl 673

• Triton 273, 287, 853

• Triton X-100^® 116, 140, 529–531, 652, 751

• Triton X-114 116, 530

• TRIZMA 45

• tRNA methyltransferase 739

• Tropomyosin 52

• Troubleshooting, for

  • cell affinity chromatography 696–677
  • chromatofocusing 413
  • chromatography on porous matrices 824
  • chromatography using Strep-tag affinity peptides 834–835
  • covalent chromatography 662–663
  • displacement chromatography 522
  • dye-ligand affinity chromatography 444–445
  • hydrophobic interaction chromatography 257–258, 266–267
  • hydroxyapatite chromatography 296–297
  • immobilized metal ion affinity chromatography 374–377
  • integral membrane proteins 548
  • ion exchange chromatography 77–78
  • lectins and glycoconjugates 634–635
  • liquid–liquid partition chromatography 501–502
  • miscellaneous biospecific affinity ligands 759–761
  • recombinant proteins 579
  • reversed-phase chromatography 194, 196
  • size exclusion chromatography 134–136

• Tryglycylglycine 688

• Trypan blue 671

• Trypsin 135

• Trypsin inhibitors 306

• Trypsinogen 141

• Tryptophan 154

• TSK 4000SW 124

• TSK Toyopearl 775

• TSK-gel G3000SW 113

• TSKgel PW series 112

• TSKgel SW type series 112

• TSK-PW-type 127

• TSK-SW-type 127

• T-suppressor cells 678–679, 688

• Tuna 304

• Tween 852–853

• Tween 20 410

• Tween 80 530

• Two-hybrid system 588

• Two-phase system 474, 532

• Two-step detergent gradients 459

• Two-step linear gradient 462

U

• Ubiquinol-cytochrome c oxidoreductase 828

• Ubiquitin 308–309

• UDP 718

• UDP-glucose dehydrogenase 430

• UDP-glucoseceramide glycosyltransferase 435

• UDP-glucuronosyltransferase 535

• Ultrafiltration 217, 623, 646, 767, 860

• Ultrasonic disintegration 561

• Ultrogel 626

• Ultrogel AcA 44 631–632

• UMP 718

• Unbound proteins 282

• Unfolding 141

• Unfolding of protein 238

• Unspecific interactions 23

• Unwanted ionic interactions 849

• Uracil 193

• Urea 38, 127, 141, 292, 314, 405, 743, 858–859

• Urtica dioica lectin 620

• UTP 718

• UV detector 93

• UV spectrometer 137

• UV/VIS-monitors 13

• UV-detection 154

• UV-monitor 12

• UV-transparency 321, 533

V

• Validation 769

• van Deemter 30, 80

• van Deemter equation 80

• van Deemter plot 30

• Vantage A, S 779

• Variable wavelength 13

• Varicella zoster virus 537

• V _e 91

• V_H domains 367, 828

• V_L domains 367

• V _o 91

• Virgil 72, 73

• Viscometers 143

• Viscosity 51, 116, 849

• Viscous solution 282

• Viscum album 624–625

• Viscum album agglutinin 624

• Vitamin B₁₂ 193

• Void volume 91, 282, 680–681

• Volatile buffers 280, 846

• Volatile ion-pair reagents 217

• Volatile mobile phases 150

W

• Wall effect 129

• Washing 454

• Water for injection 692

• Weak anion exchanger 387

• Weak ion exchanger 81

• Western blotting 842

• Wheat germ agglutinin 628, 673

• Wide bore tubing 118

• Work stations 810

X

• Xenobiotics 42

• X-ray 814

• Xylose isomerase 308

Y

• Yeast 304, 767

• Yeast cell extracts 259

• YMC–Pack Diol 101

Z

• Z-cells 154

• Z domain 563

• Zirconia 78

• Zirconium dioxide 78

• Zone widening 409

• Zorbax Bio–Series 97

• Zorbax GF-250 95

• Zorbax PSM 60 98

• Zwitter ionic detergents 405

• Zwittergent 530

• Zwitterionic 845

• Zwitterions 40