Table 4.

A summary of literature pertaining to methods adopted for isolation of various lectins from human blood (plasma/serum).

S. No.	Methods of isolation	Matrix used∗	Eluants used in adsorption chromatography	References
1. C-reactive protein
1.	Precipitation with ammonium sulphate (x2)	Not relevant	Not relevant	MacLeod and Avery (1941)
	↓
	Precipitation by dialysis against water	Not relevant	Not relevant
	Precipitation with sodium sulphate (x2)	Not relevant	Not relevant
	↓
	Precipitation by dialysis against water	Not relevant	Not relevant
2.	Precipitation with barium sulphate	Not relevant	Not relevant	Ganrot and Kindmark (1969)
	↓
	Precipitation with ammonium sulphate	Not relevant	Not relevant
	↓
	Gel adsorption	Reinagar	10 mM EDTA
3.	GF	Sephadex G - 200	Not relevant	Kushner and Somerville (1970)
4.	Density gradient centrifugation	Not relevant	Not relevant
5.	Precipitation with sodium sulphate	Not relevant	Not relevant	Siegel et al. (1974)
	↓
	GF	Sephadex G - 200	Not relevant
6.	Precipitation with ammonium sulphate (x2)	Not relevant	Not relevant	Kaplan and Volanakis (1974) Nunomura et al. (1990)
	↓
	IEC	DEAE - cellulose	1.5 M NaCl
	↓
	IEC	DEAE - cellulose	NaCl & pH gradient
7.	Precipitation with L - α – lecithin	Not relevant	Not relevant	Hokama et al. (1974)
	↓
	Precipitation by dialysis against calcium chloride	Not relevant	Not relevant
	↓
	Precipitation with chloroform	Not relevant	Not relevant
	↓
	GF	Sephadex G - 200	Not relevant
	↓
	IEC	DEAE - cellulose	NaCl gradient
8.	IEC	DEAE - cellulose (x2)	EDTA & NaCl	Johnson and Prellner (1977)
9.	AC	CPS – Sepharose	10 mM EDTA	de Beer et al. (1982)
	↓
	GF	Ultrogel AcA44	Not relevant
	↓
	IAC	Anti NHS –Sepharose	Effluent used
	↓
	GF	Sephacryl S – 300	Not relevant
10.	AC	Sepharose 4B	10 mM EDTA	de Beer and Pepys (1982)
	↓
	IAC	Anti NHS – Sepharose	Effluent used
	↓
	AC	Blue Sepharose	Effluent used
	↓
	GF	Sephacryl S – 300	Not relevant
11.	AC	CH -Sepharose 4B	2 mM EGTA	Hashimoto and Tatsumi (1989)
	↓
	HIC	Hydroxylapatite	Phosphate buffer gradient
12.	IAC	Anti CRP - Sepharose 4B	1.5 M NaCl	Nunomura et al. (1990)
	↓
	IEC	DEAE – Sephacel	500 mM NaCl
13.	AC	Sepharose 4B	Effluent used	Köttgen et al. (1992)
	↓
	AC	Phosphorylcholine - agarose	2 mM EDTA
	↓
	AC	Phosphorylcholine - agarose	1 mM phosphorylcholine
14.	AC	Phosphorylcholine - Sepharose 4B	2 mM EDTA	Culley et al. (1996)
	↓
	IEC	DEAE – cellulose	NaCl gradient
	↓
	GF	Sephacryl S – 300	Not relevant
15.	AC	Agarose beads	Effluent used	Das et al. (2004)
	↓
	AC	Phosphorylcholine - Sepharose 4B	10 mM EDTA
	↓
	AC	Phosphorylcholine - Sepharose 4B	2 mM phosphorylcholine
2. Serum amyloid protein
1.	Precipitation by dialysis against water	Not relevant	Not relevant	Binette et al. (1974)
	↓
	GF	Biogel P – 300	Not relevant
	↓
	Preparative electrophoresis	Not relevant	Not relevant
2.	AC	Sepharose 4B	50 mM sodium citrate	Pepys et al., 1977a, Pepys et al., 1977b
	↓
	GF	Ultrogel AcA 34	Not relevant
3.	Precipitation with barium chloride	Not relevant	Not relevant	Thompson and Enfield (1978)
	↓
	Precipitation with ammonium sulphate (x2)	Not relevant	Not relevant
	↓
	GF	Sephadex G – 25	Not relevant
	↓
	IEC	DEAE - Sephadex G – 25	1 mM benzamidine in sodium citrate buffer gradient
	↓
	Precipitation with ammonium sulphate	Not relevant	Not relevant
	↓
	AC	Heparin-agarose	150 mM sodium citrate
4.	AC	Sepharose 4B	25 mM EDTA	Painter et al. (1982)
	↓
	IEC	DEAE – cellulose	200 mM NaCl
5.	AC	CPS - Sepharose 4B	10 mM EDTA	Hind et al. (1984)
	↓
	GF	Ultrogel AcA44	Not relevant
	↓
	IAC	Mixture of Anti NHS - Sepharose 4B	Effluent used
	↓	& Anti SAP - Sepharose 4B
	AC	Blue Sepharose	Effluent used
	↓
	LAC	Con A – Sepharose	Effluent used
	↓
	GF	Sephacryl S – 300	Not relevant
6.	AC	Biogel A - 0.5 m	10 mM EDTA	Potempa et al. (1985)
	↓
	AC	Protein A - Sepharose CL - 4B	Effluent used
	↓
	GF	Ultrogel AcA34	Not relevant
	↓
	GF	Sephacryl S – 300	Not relevant
7.	AC	Gelatin-Sepharose 4B	Effluent used	Hamazaki (1986)
	↓
	AC	Lysine-Sepharose 4B	Effluent used
	↓
	AC	Glc - Gal - Hyl - CH Sepharose 4B	5 mM EDTA
8	AC	Sepharose 4B	5 mM EDTA	Hamazaki (1987)
	↓
	GF	TSK - GEL HW - 65S	Not relevant
9.	AC	Phosphocholine - Sepharose 4B	Effluent used	Colley et al. (1988)
	↓
	AC	Mannan - Sepharose CL - 4B	2 mM EDTA
10.	Precipitation with calcium chloride (x2)	Not relevant	Not relevant	Urbányi and Medzihradszky (1992)
	↓
	AC	Sepharose 6B	4 mM EDTA
	↓
	IEC	Sepabeads FP - DA05	NaCl gradient
11.	AC	Sepharose CL - 4B	10 mM EDTA	Danielsen et al. (1997)
	↓
	IEC	Mono – Q	NaCl gradient
12.	Precipitation with ethanol	Not relevant	Not relevant	Kilpatrick (1997b)
	↓
	Precipitation with ammonium sulphate	Not relevant	Not relevant
	↓
	AC	Emphaze - mannan (x2)	10 mM EDTA
3. H - Ficolin
1.	Isoelectric precipitation	Not relevant	Not relevant	Yae et al. (1991)
	↓
	HIC	Hydroxylapatite - Bio - Gel HTP	Phosphate buffer gradient
	↓
	Precipitation with ammonium sulphate	Not relevant	Not relevant
	↓
	GF	Sephadex G – 200	Not relevant
	↓
	Preparative electrophoresis	Not relevant	Not relevant
	↓
	LAC	Lentil lectin – agarose	200 mM α-methyl-D- mannoside
	↓
	IAC	Anti IgG - Sepharose 4B	Effluent used
2.	IAC	Anti Hakata antigen - Sepharose 4B	Effluent used	Sugimoto et al. (1998)
	↓
	IAC	Hitrap Protein G	Effluent used
	↓
	MAC	Zinc column	Glycine - HCl buffer gradient
	↓
	LAC	Lentil lectin – agarose	200 mM α -methyl - D - mannoside
3.	Precipitation with ethanol	Not relevant	Not relevant	Matsushita et al. (2002)
	↓
	Precipitation with polyethylene glycol	Not relevant	Not relevant
	↓
	AC	GlcNAc – agarose	Effluent used
	↓
	IAC	Anti H - Ficolin – Sepharose	100 mM glycine - HCl buffer
	↓
	LAC	Lentil - lectin – Sepharose	200 mM α - methyl - mannopyranoside
	↓
	IAC	Anti IgM – Sepharose	Effluent used
	↓
	AC	Protein A – Sepharose	Effluent used
	↓
	IAC	Anti MBL – Sepharose	Effluent used
	↓
	IAC	Anti L - Ficolin – Sepharose	Effluent used
4. Mannan - binding lectin
1.	AC	Mannan - Sepharose 4B (x3)	2mM EDTA	Kawasaki et al. (1983)
	↓
	GF	Sepharose CL - 6B	Not relevant
2.	AC	Sepharose 4B	Effluent used	Summerfield and Taylor (1986)
	↓
	AC	Mannan - Sepharose 4B	2 mM EDTA
3.	AC	Reacti – gel	Effluent used
	↓
	AC	Mannan - Reacti – gel	2 mM EDTA
	↓
	AC	Mannan - oxirane acrylic beads	10 mM EDTA
4.	AC	Mannan - Biogel P – 150	10 mM EDTA	Taylor and Summerfield (1987)
	↓
	GF	Sepharose CL - 6B	Not relevant
5.	AC	Phosphocholine - Sepharose CL - 4B	Effluent used	Colley et al. (1988)
	↓
	AC	Mannan - Sepharose CL - 4B	2 mM EDTA
	↓
	AC	Mannan - Sepharose CL - 4B	50 mM mannose
6.	GF	Sephacryl - S300	Not relevant	Super et al. (1989)
	↓
	AC	Mannan – Sepharose	5 mM EDTA
	↓
	IAC	Anti IgM – Sepharose	Effluent used
	↓
	IEC	Mono – Q	1M NaCl
7.	AC	Mannan - Sepharose	5 mM EDTA
	↓
	AC	Mannan – Sepharose	Mannose
	↓
	GF	Superose 6	Not relevant
	↓
	IEC	Mono – Q	1 M NaCl
	↓
	IAC	Anti IgM – Sepharose	Effluent used
8.	AC	Mannan - Sepharose 4B	2 mM EDTA	Kuhlman et al. (1989)
	↓
	AC	Mannan - Sepharose 4B	50 mM mannose
9.	AC	Mannan - Sepharose 4B	10 mM EDTA	Lu et al. (1990)
	↓
	AC	Mannan - Sepharose 4B	50 mM mannose
	↓
	GF	Superose 6 (HR10/30)	Not relevant
	↓
	IEC	Mono - Q (HR5/5)	NaCl gradient
	↓
	IAC	Anti IgM - Sepharose	Effluent used
10.	AC	Mannose - Sepharose 6B	10 mM EDTA	Kyogashima et al. (1990)
	↓
	AC	Sepharose 6B	10 mM mannose
11.	Precipitation with polyethylene glycol	Not relevant	Not relevant	Matsushita and Fujita (1992)
	↓
	AC	Mannan - Sepharose 4B	300 mM mannose
	↓
	IAC	Anti IgM - Sepharose 4B	Effluent used
	↓
	IAC	Anti MBP - Sepharose 4B (x2)	100 mM glycine - HCl buffer
12.	AC	Mannose - Sepharose 6B	10 mM EDTA	Terai et al. (1993)
	↓
	AC	Sepharose 6B	50 mM mannose
	↓
	GF	Superose 6	Not relevant
	↓
	IEC	Mono – Q	NaCl gradient
13.	Precipitation with polyethylene glycol	Not relevant	Not relevant	Tan et al. (1996)
	↓
	AC	Mannose - Sepharose 4B	10 mM EDTA
	↓
	AC	Maltose - Sepharose 4B	100 mM N - acetylglucosamine
	↓
	IEC	Mono - Q (HR5/5)	NaCl gradient
	↓
	AC	Mannose-Sepharose 4B	10 mM EDTA
	↓
	GF	Superose 6	Not relevant
14.	Precipitation with ethanol	Not relevant	Not relevant	Kilpatrick (1997a)
	↓
	Precipitation with ammonium sulphate	Not relevant	Not relevant
	↓
	AC	Emphaze – mannan	10 mM EDTA
	↓
	AC	Emphaze – mannan	100 mM mannose
15.	AC	Mannan - Sepharose 4B (x2)	20 mM EDTA	Suankratay et al. (1998)
	↓
	AC	Protein A – Sepharose	Effluent used
	↓
	AC	Anti IgM – Sepharose	Effluent used
16.	AC	Mannan - Sepharose 4B (x2)	20 mM EDTA	Saifuddin et al. (2000)
	↓
	AC	Protein G – Sepharose	Effluent used
	↓
	IAC	Anti IgM – Sepharose	Effluent used
17.	Precipitation with polyethylene glycol	Not relevant	Not relevant	Matsushita et al. (2000)
	↓
	AC	GlcNAc – agarose	300 mM mannose
	↓
	IAC	Anti MBL - Sepharose 4B	100 mM glycine - HCl buffer
18.	Precipitation with polyethylene glycol	Not relevant	Not relevant	Muto et al. (2001)
	↓
	AC	Mannan – agarose	10 mM EDTA
	↓
	AC	Mannan – agarose	50 mM mannose
	↓
	GF	Sephacryl S – 300	Not relevant
	↓
	IAC	Anti IgM – Sepharose	Effluent used
	↓
	AC	Protein G – Sepharose	Effluent used
19.	Precipitation with ethanol	Not relevant	Not relevant	Neth et al. (2002)
	↓
	Precipitation with ammonium sulphate	Not relevant	Not relevant
	↓
	AC	Mannan – agarose	10 mM EDTA
	↓
	AC	Mannan – agarose	100 mM mannose
20.	AC	Mannose - Sepharose 4B	10 mM EDTA	Butler et al. (2002)
	↓ ↓
	AC	Maltose - Sepharose 4B	100 mM N - acetylglucosamine
	↓
	GF	Sephacryl S – 300	Not relevant
	↓
	IAC	Anti α2 - macroglobulin - Sepharose 4B	Effluent used
21.	Precipitation with ethanol	Not relevant	Not relevant	Matsushita et al. (2002)
	↓
	Precipitation with polyethylene glycol	Not relevant	Not relevant
	↓
	AC	GlcNAc – agarose	300 mM mannose
	↓
	IAC	Anti MBL - Sepharose 4B	100 mM glycine - HCl buffer
22.	Precipitation with ethanol	Not relevant	Not relevant	Valdimarsson et al. (2003)
	↓
	AC	Agarose	30 mM mannose
	↓
	IEC	Q-Sepharose	NaCl
	↓
	GF	Superose 6	Not relevant
23.	AC	Sepharose CL - 4B	30 mM mannose	Laursen, 2003
	↓
	IEC	Q – Sepharose	NaCl
	↓
	GF	Superose 6	Not relevant
24.	Precipitation with polyethylene glycol	Not relevant	Not relevant	Ma et al. (2004)
	↓
	AC	Peptidoglycan - Sepharose 4B	300 mM mannose
	↓
	AC	Protein A - Sepharose CL - 4B	Effluent used
	↓
	IAC	Anti IgM - Sepharose 4B	Effluent used
5. Tetranectin
1.	Precipitation with barium citrate	Not relevant	Not relevant	Clemmensen et al. (1986)
	↓
	AC	Lysine - Sepharose 4B	Effluent used
	↓
	Precipitation with ammonium sulphate	Not relevant	Not relevant
	↓
	AC	Plasminogen - Sepharose 4B	1 mM tranexamic acid
	↓
	IEC	DEAE - Sepharose CL - 6B	NaCl gradient
	↓
	GF	Ultrogel AcA34	Not relevant
2.	Cryoprecipitate depletion	Not relevant	Not relevant	Fuhlendorff et al. (1987)
	↓
	IAC	Antitetranectin - Sepharose 4B	3 M MgCl2
	↓
	IAC	Antihuman plasma protein column	Effluent used
	↓
	GF	Ultrogel AcA34	Not relevant
3.	AC	Hitrap Heparin - Sepharose	Phosphate buffer gradient	Thougaard et al. (2001)
6. L - Ficolin
1.	Polyethylene glycol precipitation	Not relevant	Not relevant	Matsushita et al. (1996)
	↓
	AC	Mannan - Sepharose 4B	150 mM N - acetylglucosamine
	↓
	IEC	Mono – Q	NaCl gradient
2.	AC	Sepharose 4B	Effluent used	Le et al. (1997)
	↓
	AC	GlcNAc - Sepharose 4B	100 mM N - acetylglucosamine
	↓
	IAC	Q - Sepharose 4B	Effluent used
	↓
	IEC	Mono – Q	NaCl gradient
	↓
	AC	Tris - blocked CNBr - activated Sepharose 4B	100 mM N - acetylglucosamine
3.	AC	Sepharose 4B	Effluent used	Le et al. (1998)
	↓
	AC	GlcNAc - Sepharose 4B	200 mM N - acetylglucosamine
	↓
	IEC	Mono - Q (x2)	NaCl gradient
	↓
	AC	Tris - blocked CNBr - activated Sepharose 4B	200 mM N - acetylglucosamine
4.	Precipitation with polyethylene glycol	Not relevant	Not relevant	Matsushita et al.,2000
	↓
	AC	GlcNAc – agarose	150 mM N - acetylglucosamine
	↓
	IEC	Mono – Q	NaCl gradient
	↓
	AC	Anti MBL - Sepharose 4B	Effluent used
5.	Precipitation with ethanol	Not relevant	Not relevant	Matsushita et al. (2002)
	↓
	Precipitation with polyethylene glycol	Not relevant	Not relevant
	↓
	AC	GlcNAc – agarose	150 mM N - acetylglucosamine
	↓
	IEC	Mono – Q	NaCl gradient
6.	Precipitation with ethanol	Not relevant	Not relevant	Cseh et al. (2002)
	↓
	Precipitation with polyethylene glycol	Not relevant	Not relevant
	↓
	AC	GlcNAc – agarose	300 mM N - acetylglucosamine
	↓
	AC	Asialofetuin - Sepharose (x2)	300 mM N - acetylglucosamine
	↓
	IAC	Anti MBL - Sepharose 4B	Effluent used
	↓
	IAC	Anti H- ficolin - Sepharose 4B	Effluent used
7.	Polyethylene glycol precipitation	Not relevant	Not relevant	Ma et al. (2004)
	↓
	AC	1, 3 -β-D-glucan-Toyopearl	300 mM N - acetylglucosamine
8.	Polyethylene glycol precipitation	Not relevant	Not relevant	Krarup et al. (2004)
	↓
	AC	N - acetylcysteine - Sepharose CL - 4B	Lower ionic strength buffer
	↓
	IEC	Mono – Q	NaCl gradient

Number given in parenthesis indicates the successive repetition of the same method employed.

Abbreviations used: AC = Affinity chromatography; CPS = Capsular polysaccharide; Con A = Concanavalin A; CNBr = Cyanogen bromide; CRP = C-reactive protein; DEAE = Diethylaminoethyl; EDTA = Ethylenediaminetetraacetic acid disodium salt; EGTA = Ethylene glycol -bis-(β - aminoethylether) N, N, Ń, Ń - tetraacetic acid; GF = Gel filtration; Glc-Gal-Hyl = 2-O-α-D-glucopyranosyl-O-β-D- galactopyranosyl hydroxylysine; HIC = Hydrophobic interaction chromatography; IAC = Immuno - affinity chromatography; IEC = Ion exchange chromatography; IgG = Immunoglobulin G; Immunoglobulin M = IgM; LAC = Lectin affinity chromatography; MAC = Metal affinity chromatography; MBL = Mannan-binding lectin; MBP = Mannan-binding protein; NHS = Normal human serum; SAP = Serum amyloid protein.

^∗The gel type of the matrix is given as reported by the investigators.