KEY RESOURCES TABLE
| REAGENT or RESOURCE | SOURCE | IDENTIFIER |
|---|---|---|
| Antibodies | ||
| Anti-human CD32 | BD Biosciences | Cat# 551900; RRID: AB_394290 |
| Anti-human CD16 | BD Biosciences | Cat# 550383; RRID: AB_393649 |
| Anti-human CD23 | BD Biosciences | Cat# 555707; RRID: AB_396053 |
| Anti-human CD19-BV711 | BioLegend | Cat# 302246; RRID: AB_2562065 |
| Anti-human CD27-PE-Cy7 | eBioscience | Cat# 25-0271-82; RRID: AB_1724035 |
| Anti-human CD14-FITC | BD Biosciences | Cat# 557153; RRID: AB_396589 |
| Anti-human CD3-FITC | BD Biosciences | Cat# 556611; RRID: AB_396484 |
| Anti-human CD20-Alexa Fluor 700 | BioLegend | Cat# 302322; RRID: AB_493753 |
| Anti-human IgD-PerCP-Cy5.5 | eBioscience | Cat# 46-9868-42; RRID: AB_2573920 |
| Anti-human IgM-BV605 | BioLegend | Cat# 314524; RRID:AB_2562374 |
| Anti-human IgG-APC | BD Biosciences | Cat# 550931; RRID: AB_398478 |
| Anti-human IgG-FITC | Jackson ImmunoResearch | Cat# 109-095-008; RRID: AB_2337651 |
| Anti-mouse CD19-BUV395 | BD Biosciences | Cat# 563557; RRID: AB_2722495 |
| Anti-mouse CD3-BV711 | BioLegend | Cat# 100349; RRID: AB_2565841 |
| Anti-mouse Ly6G-BV711 | BioLegend | Cat# 127643; RRID: AB_2565971 |
| Anti-mouse F4/80-BV711 | BioLegend | Cat# 123147; RRID: AB_2564588 |
| Anti-mouse B220-BV786 | BD Biosciences | Cat# 563894; RRID: AB_2738472 |
| Anti-mouse GL7-PerCP-Cy5.5 | BioLegend | Cat# 144609; RRID: AB_2562978 |
| Anti-mouse CD45.2-Alexa Fluor 647 | BioLegend | Cat# 109818; RRID: AB_492870 |
| Anti-mouse CD38-Alexa Fluor 700 | eBioscience | Cat# 56-0381-82; RRID: AB_657740 |
| Anti-mouse CD45.1-PE-eFluor 610 | eBioscience | Cat# 61-0453-82; RRID: AB_2574560 |
| Anti-mouse Bcl6-FITC | BioLegend | Cat# 358513; RRID: AB_2860942 |
| HRP-conjugated goat anti-mouse IgG | BioLegend | Cat# 405306; RRID: AB_315009 |
| Biological Samples | ||
| PMBC from HIV-uninfected donors (Establishing Immunologic Assays for Determining HIV-1 Prevention and Control) | This paper | N/A |
| Chemicals, peptides, and recombinant proteins | ||
| Streptavidin-BV421 | BioLegend | Cat# 405226 |
| Fixable Viability dye-eFluor 506 | eBioscience | Cat# 65-0866-14 |
| Protein A agarose resin | Thermo Scientific | Cat# 20333 |
| IgG elution buffer | Thermo Scientific | Cat# 21004 |
| Endoproteinase LysC | New England BioLabs | Cat# P8109S |
| His60 Ni superflow resin | Takara | Cat# 635664 |
| HiLoad 16/600 Superdex 200 pg column | GE Healthcare | Cat# 28-9893-35 |
| Ni-NTA agarose resin | QIAGEN | Cat# 30230 |
| ENrich SEC 650 10 × 300 column | Biorad | Cat# 7801650 |
| DyLight 550 NHS ester | Thermo Scientific | Cat# 62262 |
| DyLight 650 NHS ester | Thermo Scientific | Cat# 62266 |
| Zenon Allophycocyanin human IgG labeling kit | Thermo Scientific | Cat# Z25451 |
| Zenon R-Phycoerythrin human IgG labeling kit | Thermo Scientific | Cat# Z25455 |
| Fluo-4 direct calcium indicator | Thermo Scientific | Cat# F10471 |
| Ionomycin | Millipore-Sigma | Cat# I9657 |
| SureBlue Reserve TMB Peroxidase Substrate | Seracare KPL | Cat# 5120-0080 |
| CellTrace Violet (CTV) | Thermo Scientific | Cat# C34557 |
| Sigma Adjuvant System | Sigma | Cat# S6322 |
| FoxP3/Transcription factor Fix and Perm buffer | eBioscience | Cat# 00-5523-00 |
| RBC lysing buffer | Sigma | Cat# R7757 |
| QuantumPlex M Carobxyl magnetic beads | Bangs Laboratories | Cat# 250 |
| Mouse Ig-Primer set | Millipore-Sigma | Cat# 69831 |
| GeneJuice | Millipore-Sigma | Cat# 70967 |
| 293 Freestyle media | Thermo Scientific | Cat# 12-338-018 |
| 293 Free transfection reagent | Millipore-Sigma | Cat# 72181 |
| Critical commercial assays | ||
| Mouse B cell isolation kit | StemCell Technologies, Inc. | Cat# 19854 |
| Human B cell enrichment kit | StemCell Technologies, Inc. | Cat# 19054 |
| iScript | Bio-Rad | Cat# 1708891 |
| Deposited data | ||
| ai-mAb VH and VL sequences | This paper | GenBank: MT561048 - MT561059 |
| iv1-iGL-VRC01 complex | This paper | PDB: 7JLK |
| iv4-iGL-VRC01 complex | This paper | PDB: 6XOC |
| Iv9-iGL-VRC01 complex | This paper | PDB: 7JLN |
| Iv12-iGL-12A21 complex | This paper | PDB: 6VRQ |
| Experimental models: Cell lines | ||
| 293-6E | National Research Council, Canada | N/A |
| HEK293T | ATCC | Cat#C RL-3216 |
| DG-75 B cells | ATCC | Cat# CRL-2625 |
| Experimental models: Organisms/strains | ||
| iGL-VRC01 HC homozygous mice | This paper | N/A |
| iGL-VRC01 LC homozygous mice | This paper | N/A |
| iGL-VRC01 HC+LC homozygous mice | This paper | N/A |
| B6.SJL-Ptprca Pepcb/BoyJ | The Jackson Laboratory | 002014 |
| C57BL/6J | The Jackson Laboratory | 000664 |
| Oligonucleotides | ||
| Primers for antibody nested PCR and sequencing | Tiller et al., 2008 | N/A |
| Spycatcher (AA 24-121) | Zakeri et al., 2012 | GenBank: AFD50637.1 |
| Spytag: GGSGGSGHHHHHHGSAHIVMVDAYKPTK | Zakeri et al., 2012 | N/A |
| Recombinant DNA | ||
| pTT3-iv4-Fab-HC-SC | This paper | N/A |
| pTT3-iv9-Fab-HC-ST | This paper | N/A |
| pTT3-iv1-scFv | This paper | N/A |
| pTT3-iGL-VRC01-HC-Fab | This paper | N/A |
| pTT3-iv4 LC | This paper | N/A |
| pTT3-iv9 LC | This paper | N/A |
| pTT3-eOD-GT8 | Jardine et al., 2015 | N/A |
| pTT3-eOD-GT8-KO | Dosenovic et al., 2019 | N/A |
| pRRL.EuB29.VRC01.BCR.GFP.WPRE | McGuire et al., 2014a | N/A |
| psPAX2 | Addgene | Cat# 12260 |
| pMD2.G | Addgene | Cat# 12259 |
| Software and algorithms | ||
| Flow Jo, version 9.9.4 | Tree Star | https://www.flowjo.com |
| Prism, version 8.4.1 | GraphPad Software | https://www.graphpad.com |
| ForteBio data analysis software | ForteBio | N/A |
| HKL2000 | Otwinowski and Minor, 1997 | https://www.hkl-xray.com/how-reference-hkl-hkl-2000-and-hkl-3000 |
| Phaser | McCoy et al., 2007 | https://www.phenix-online.org/documentation/reference/phaser.html |
| Phenix.refine | Afonine et al., 2012 | https://www.phenix-online.org/documentation/reference/refinement.html |
| Coot | Emsley et al., 2010 | RRID:SCR_014222 |
| Pymol | Delano, 2002 | https://pymol.org/2/ |
| Other | ||
| SpectraMax M2 plate reader | Molecular Devices | https://www.moleculardevices.com |
| BioTek 405 select microplate washer | BioTek | https://www.biotek.com |
| FACS Aria II | Becton, Dickinson and Company | N/A |
| Symphony X50 flow cytometer | Becton, Dickinson and Company | N/A |
| IQue3 high throughput flow cytometer | Intellicyt | N/A |
| Octet RED96e | ForteBio | N/A |
| Anti-mouse IgG Fc capture (AMC) biosensors | ForteBio | Cat# 18-5088 |
| Anti-human IgG Fc capture (AHC) biosensors | ForteBio | Cat# 18-5060 |
| Streptavidin biosensors | ForteBio | Cat# 18-5019 |
| Rock Imager | Formulatrix | N/A |