Characterization of Anti-HIV Antibodies

Human HIV-1 IgG antibodies used as reference controls are as follows: anti-V3^crown 10-188 (Mouquet et al., 2011 (link)); bNAbs 2G12 (Trkola et al., 1996 (link)), anti-CD4bs 3BNC117 (Scheid et al., 2011 (link)), anti-V3-glycan PGT121, PGT135, 10-1074, and BG8/BG18 (Freund et al., 2017 (link); Mouquet et al., 2012 (link); Walker et al., 2011 (link)); and anti-silent face SF12 (Schoofs et al., 2019 (link)). Non–HIV-1 antibodies include polyreactive and nonpolyreactive antibody ED38 (Meffre et al., 2004 (link)) and mGO53 (Wardemann et al., 2003 (link)), respectively. 10-1074 and mGO53 IgA monoclonal antibodies were also generated (Lorin et al., 2017 (link)). Recombinant IgG and IgA antibodies were produced by cotransfection of Freestyle 293-F cells (Thermo Fisher Scientific) using PEI precipitation method as previously described (Lorin and Mouquet, 2015 (link); Tiller et al., 2008 (link)) and purified by affinity chromatography using protein G Sepharose 4 fast flow beads (GE Healthcare) and peptide M-coupled agarose beads (InvivoGen), respectively. For competition ELISA experiments, purified antibodies were biotinylated using the EZ-Link Sulfo-NHS-Biotin kit (Thermo Fisher Scientific).

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Lorin V., Fernández I., Masse-Ranson G., Bouvin-Pley M., Molinos-Albert L.M., Planchais C., Hieu T., Péhau-Arnaudet G., Hrebík D., Girelli-Zubani G., Fiquet O., Guivel-Benhassine F., Sanders R.W., Walker B.D., Schwartz O., Scheid J.F., Dimitrov J.D., Plevka P., Braibant M., Seaman M.S., Bontems F., Di Santo J.P., Rey F.A, & Mouquet H. (2022). Epitope convergence of broadly HIV-1 neutralizing IgA and IgG antibody lineages in a viremic controller. The Journal of Experimental Medicine, 219(3), e20212045.

Publication 2022

Freestyle 293-F cells protein G Sepharose 4 fast flow beads peptide M-coupled agarose beads EZ-Link Sulfo-NHS-Biotin kit

Affinity chromatography Agarose Antibodies Antibody Bnabs Cells Elisa Face Glycan Hiv 1 Hiv antibodies Human Iga antibodies Igg antibodies Monoclonal antibodies Peptide m Protein g Sulfo nhs biotin Walker

Corresponding Organization : Inserm

Other organizations : Sorbonne Paris Cité, Université Paris Cité, Department of Virology, Université de Tours, Central European Institute of Technology, Central European Institute of Technology – Masaryk University, University of Amsterdam, Cornell University, Amsterdam University Medical Centers, Ragon Institute of MGH, MIT and Harvard, Harvard University, Massachusetts General Hospital, Rockefeller University, Centre de Recherche des Cordeliers, Sorbonne Université, Beth Israel Deaconess Medical Center, Institut de Chimie des Substances Naturelles, Université Paris-Saclay

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Variable analysis

independent variables

Anti-V3^crown 10-188
BNAbs 2G12
Anti-CD4bs 3BNC117
Anti-V3-glycan PGT121, PGT135, 10-1074, and BG8/BG18
Anti-silent face SF12
Polyreactive and nonpolyreactive antibody ED38
MGO53
10-1074 and mGO53 IgA monoclonal antibodies

dependent variables

Not explicitly mentioned

control variables

Recombinant IgG and IgA antibodies were produced by cotransfection of Freestyle 293-F cells (Thermo Fisher Scientific) using PEI precipitation method as previously described (Lorin and Mouquet, 2015; Tiller et al., 2008) and purified by affinity chromatography using protein G Sepharose 4 fast flow beads (GE Healthcare) and peptide M-coupled agarose beads (InvivoGen), respectively.
For competition ELISA experiments, purified antibodies were biotinylated using the EZ-Link Sulfo-NHS-Biotin kit (Thermo Fisher Scientific).

controls

Positive controls: None specified
Negative controls: None specified

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