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Comparing methods for immobilizing HIV-1 SOSIPs in ELISAs that evaluate antibody binding

Dam, Kim-Marie A. and Mutia, Patricia S. and Bjorkman, Pamela J. (2022) Comparing methods for immobilizing HIV-1 SOSIPs in ELISAs that evaluate antibody binding. Scientific Reports, 12 . Art. No. 11172. ISSN 2045-2322. PMCID PMC9247892. doi:10.1038/s41598-022-15506-x. https://resolver.caltech.edu/CaltechAUTHORS:20220707-978213000

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Abstract

Enzyme-linked immunosorbent assays (ELISAs) are used to evaluate binding of broadly neutralizing antibodies (bNAbs) and polyclonal sera to native-like HIV-1 Env SOSIPs. Methods for immobilizing SOSIPs on plates differ, which can lead to variable or, in some cases, misleading results. Three methods used to immobilize SOSIPs were compared to determine how antigen immobilization methods affect Env conformation and ELISA results. HIV-1 SOSIPs were directly coated on polystyrene plates, captured by a monoclonal antibody against a C-terminal affinity tag, or randomly biotinylated and coated on a streptavidin plate. Binding of bNAbs with known epitopes were compared for each immobilization method. Binding of bNAbs targeting the V1V2, V3, CD4 binding site, and gp120/gp41 interface was comparable for all antigen immobilization methods. However, directly coated HIV-1 SOSIP ELISAs showed detectable binding of 17b, a CD4-induced antibody that binds a V3 epitope that is concealed on closed prefusion Env trimers in the absence of added CD4, whereas antibody-immobilized and randomly biotinylated Env-coated ELISAs did not show detectable binding of 17b in the absence of CD4. We conclude direct coating of HIV-1 SOSIPs on ELISA plates can result in exposure of CD4-induced antibody epitopes, suggesting disruption of Env structure and exposure of epitopes that are hidden in the closed, prefusion trimer.


Item Type:Article
Related URLs:
URLURL TypeDescription
https://doi.org/10.1038/s41598-022-15506-xDOIArticle
http://www.ncbi.nlm.nih.gov/pmc/articles/pmc9247892/PubMed CentralArticle
ORCID:
AuthorORCID
Dam, Kim-Marie A.0000-0002-1416-4757
Bjorkman, Pamela J.0000-0002-2277-3990
Additional Information:© Te Author(s) 2022��. This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. Received 18 May 2022. Accepted 24 June 2022. Published 01 July 2022. We thank J. Vielmetter, P. Hoffman, the Protein Expression Center in the Beckman Institute at Caltech for expression assistance, and J.E. Robinson (Tulane University) for the JR-52 antibody. This work was supported by the National Institute of Allergy and Infectious Diseases (NIAID) Grant HIVRAD P01 AI100148 (to P.J.B.) and the Bill and Melinda Gates Foundation Collaboration for AIDS Vaccine Discovery (CAVD) grant INV-002143 (P.J.B.). Contributions. K.A.D. and P.J.B. designed the research. K.A.D. and P.S.M. performed experiments and K.A.D. and P.J.B. analyzed results. K.A.D. and P.J.B. wrote the manuscript with input from co-authors. Data availability. The datasets generated during and analyzed in this study are available from the corresponding author on reasonable request. The authors declare no competing interests.
Funders:
Funding AgencyGrant Number
NIHP01 AI100148
Bill and Melinda Gates FoundationINV-002143
PubMed Central ID:PMC9247892
DOI:10.1038/s41598-022-15506-x
Record Number:CaltechAUTHORS:20220707-978213000
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20220707-978213000
Official Citation:Dam, KM.A., Mutia, P.S. & Bjorkman, P.J. Comparing methods for immobilizing HIV-1 SOSIPs in ELISAs that evaluate antibody binding. Sci Rep 12, 11172 (2022). https://doi.org/10.1038/s41598-022-15506-x
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:115424
Collection:CaltechAUTHORS
Deposited By: George Porter
Deposited On:08 Jul 2022 23:10
Last Modified:25 Jul 2022 23:13

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