Hidden lineage complexity of glycan-dependent HIV-1 broadly neutralizing antibodies uncovered by digital panning and native-like gp140 trimer

Linling He, Xiaohe Lin, Natalia deVal, Karen L. Saye-Francisco, Colin J. Mann, Ryan Augst, Charles D. Morris, Parisa Azadnia, Bin Zhou, Devin Sok, Gabriel Ozorowski, Andrew B. Ward, Dennis R. Burton and Jiang Zhu
Frontiers in Immunology  2017 8
@ARTICLE {, author = "He, L., Lin, X., de Val, N., Saye-Francisco, K. L., Mann, C. J., Augst, R., Morris, C. D., Azadnia, P., Zhou, B., Sok, D., Ozorowski, G., Ward, A. B., et al.", title = "Hidden Lineage Complexity of Glycan-Dependent HIV-1 Broadly Neutralizing Antibodies Uncovered by Digital Panning and Native-Like gp140 Trimer", journal = "Front. Immunol., 24 August 2017", year = "2017"}


Germline precursors and intermediates of broadly neutralizing antibodies (bNAbs) are essential to the understanding of humoral response to HIV-1 infection and B-cell lineage vaccine design. Using a native-like gp140 trimer probe, we examined antibody libraries constructed from donor-17, the source of glycan-dependent PGT121-class bNAbs recognizing the N332 supersite on the HIV-1 envelope glycoprotein. To facilitate this analysis, a digital panning method was devised that combines biopanning of phage-displayed antibody libraries, 900 bp long-read next-generation sequencing, and heavy/light (H/L)-paired antibodyomics. In addition to single-chain variable fragments resembling the wild-type bNAbs, digital panning identified variants of PGT124 (a member of the PGT121 class) with a unique insertion in the heavy chain complementarity-determining region 1, as well as intermediates of PGT124 exhibiting notable affinity for the native-like trimer and broad HIV-1 neutralization. In a competition assay, these bNAb intermediates could effectively compete with mouse sera induced by a scaffolded BG505 gp140.681 trimer for the N332 supersite. Our study thus reveals previously unrecognized lineage complexity of the PGT121-class bNAbs and provides an array of library-derived bNAb intermediates for evaluation of immunogens containing the N332 supersite. Digital panning may prove to be a valuable tool in future studies of bNAb diversity and lineage development.

Technology Platform

Digital Panning

Research Topics

Antibody Discovery

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