Fig_3.tif (2.01 MB)
Serum recognition of the native trimer and effects of a key mutation in the CD4 binding loop.
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posted on 2015-05-29, 03:58 authored by Ema T. Crooks, Tommy Tong, Bimal Chakrabarti, Kristin Narayan, Ivelin S. Georgiev, Sergey Menis, Xiaoxing Huang, Daniel Kulp, Keiko Osawa, Janelle Muranaka, Guillaume Stewart-Jones, Joanne Destefano, Sijy O’Dell, Celia LaBranche, James E. Robinson, David C. Montefiori, Krisha McKee, Sean X. Du, Nicole Doria-Rose, Peter D. Kwong, John R. Mascola, Ping Zhu, William R. Schief, Richard T. Wyatt, Robert G. Whalen, James M. BinleyMAbs and serum binding to the native Env trimer was assessed in BN-PAGE shift assays. Briefly, mAbs (30μg/ml) or sera (1:2 dilution) were incubated with protease-digested 2nd generation A) JR-FL SOS E168K VLPs or B) JR-FL SOS E168K+D368R VLPs. The VLPs were then washed and lysed, and Env was resolved by BN-PAGE-Western blot. Ferritin was used as a molecular weight marker. The unliganded trimer is indicated by a cartoon. The abbreviation “UND VLP” (immunogen for rabbit 617) refers to “undigested VLPs”.
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nAb titersbioinformatics analysisCD 4 Binding Site ElicitingNeutralizing Antibodies Bindglycan defensesclade B tier 2DNA trimer sera8 rabbitstier 2 phenotypepglimits access3 seraancvaccine researchresidue 197CD 4 binding sitefuture developmenttier 2 breadth20 rabbitsN 197 glycanglycan fenceneutralizing seraEnv trimerhivneutralizing antibody vaccinequaternary epitopestrimer VLP seratier 2 neutralizing antibodiesneutralizing DNA trimer serumdata show
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