Finton KA, Friend D, Jaffe J, Gewe M, Holmes MA, Larman HB, Stuart A, Larimore K, Greenberg PD, Elledge SJ, Stamatatos L, Strong RK. *PLoS Pathogens* · September 25, 2014 · DOI: [10.1371/journal.ppat.1004403](https://doi.org/10.1371/journal.ppat.1004403) How to cite ### AMA Finton KA, Friend D, Jaffe J, et al. Ontogeny of recognition specificity and functionality for the broadly neutralizing anti-HIV antibody 4E10. *PLoS Pathog*. 2014;10(9):e1004403. doi:10.1371/journal.ppat.1004403 ### APA ``` Finton, K. A., Friend, D., Jaffe, J., Gewe, M., Holmes, M. A., Larman, H. B., Stuart, A., Larimore, K., Greenberg, P. D., Elledge, S. J., Stamatatos, L., & Strong, R. K. (2014). Ontogeny of recognition specificity and functionality for the broadly neutralizing anti-HIV antibody 4E10. *PLoS Pathogens*, 10(9), e1004403. https://doi.org/10.1371/journal.ppat.1004403 ``` ### BibTeX ``` @article{Finton20144E10Ontogeny, author = {Finton, Kathryn A. and Friend, Della and Jaffe, James and Gewe, Mesfin and Holmes, Margaret A. and Larman, H. Benjamin and Stuart, Andrew and Larimore, Kevin and Greenberg, Philip D. and Elledge, Stephen J. and Stamatatos, Leonidas and Strong, Roland K.}, title = {Ontogeny of recognition specificity and functionality for the broadly neutralizing anti-HIV antibody 4E10}, journal = {PLoS Pathogens}, volume = {10}, number = {9}, pages = {e1004403}, year = {2014}, doi = {10.1371/journal.ppat.1004403} } ``` Finton and colleagues reconstructed an ensemble of **twelve plausible germline-encoded precursors (GEPs)** of the broadly neutralizing anti-HIV antibody 4E10, then compared their structures, biophysics, binding kinetics, and polyspecificity head-to-head with mature 4E10 using thermal-stability assays, surface plasmon resonance, crystallography of three GEPs alone and bound to engineered scaffolds, and PhIP-Seq profiling against a phage-displayed human peptidome. The result overturns the textbook account of antibody affinity maturation: 4E10 gained roughly 100-fold affinity primarily by improving off-rates against a small, conserved epitope — while becoming *less* thermostable, retaining combining-site flexibility, and acquiring autoreactivity its precursors did not have. [ Read publication at PLoS Pathogens ](https://doi.org/10.1371/journal.ppat.1004403) In this publication: - **12 germline-encoded precursors** of 4E10 were inferred via phylogenetic reconstruction across multiple V-gene-segment-assignment tools; eight were expressed as single-chain Fvs and characterized in parallel with mature 4E10 (which carries 33–35 somatic mutations). - **Mature 4E10 is markedly less thermostable than its precursors** (Tm = 52.8°C vs GEP Tm = 64.2–67.0°C) — the opposite of the rigidify-and-stabilize trajectory predicted by the consensus model of affinity maturation. - **~100-fold affinity gain came almost entirely from off-rate improvement:** 4E10 bound the engineered T117 scaffold at KD = 28 pM versus 1.8–4.0 nM for GEPs, with on-rates largely unchanged across maturation. - **Polyspecificity did not narrow during maturation** — PhIP-Seq against a human peptidome showed both 4E10 and its GEPs had limited polyspecificity, but recognized *different* off-target peptide sets, with mature 4E10 acquiring autoreactivity that the inferred precursors lacked. 4E10 is one of a small set of human antibodies that can broadly neutralize HIV by binding a conserved stretch of the viral envelope protein gp41 called the membrane-proximal external region (MPER). HIV vaccine designers have spent more than a decade trying to figure out how to make a vaccine that would re-elicit 4E10-like antibodies in a healthy person, because the natural process that produced 4E10 in an HIV-infected donor takes years of chronic antigen exposure and dozens of mutations. To do that rationally, the field needs to know what the precursor B-cell receptor that started 4E10's maturation actually looked like, and which mutations along the way did the load-bearing work. The authors used phylogenetic inference across multiple V-gene-segment-assignment tools to predict an ensemble of twelve plausible germline-encoded precursors (GEPs) of 4E10, then expressed eight of them as single-chain variable fragments (Fvs) and characterized them in parallel with mature 4E10. They measured thermostability by circular dichroism, neutralization potency against three HIV-1 isolates in TZM-bl assays, binding to soluble Env trimers (gp1403) and four engineered epitope-scaffold (ES) immunogens by surface plasmon resonance, and crystal structures of three GEPs alone and bound to the T117 scaffold. To compare polyspecificity and autoreactivity between mature 4E10 and its precursors, they applied phage-immunoprecipitation sequencing (PhIP-Seq) against a phage-displayed human peptidome. The findings cut against the consensus model of affinity maturation in three specific ways. First, 4E10's thermostability (Tm = 52.8°C) was markedly worse than its predicted GEPs (Tm = 64.2 to 67.0°C). Second, both 4E10 and its GEPs sampled extensive heavy-chain CDR conformer ensembles in crystal structures; maturation did not rigidify the combining site as the standard model predicts. Third, the polyspecificity expected to narrow during maturation did not narrow — PhIP-Seq showed both 4E10 and its GEPs had limited polyspecificity, but they recognized *different* off-target peptide sets, with 4E10 acquiring its characteristic autoreactivity that the GEPs lacked. The roughly 100-fold affinity gain to the engineered T117 scaffold came almost entirely through faster on-rates becoming slower off-rates, rather than through the entropy gains the consensus model predicts. For HIV vaccine designers using germline-targeting strategies, the work argues for explicit design of unconventional maturation pathways — sequential immunogens that select for autoreactivity tolerance, off-rate optimization, and continued combining-site flexibility, rather than the standard rigidify-and-narrow trajectory. [← Back to all resources](/) ## Related resources [![](https://cdn.sanity.io/images/vbcbvda6/production/d789ad5a7e0a909e5a212ab947adc6aefa3d99c8-1200x750.webp)White Paper ### Unbiased analysis of antibody responses to all human viruses using Infinity Bio's VirSIGHT library A peer-reviewed methodology white paper for VirSIGHT, Infinity Bio's unbiased antibody reactome assay covering 500+ huma… Read more ](/resources/unbiased-analysis-of-antibody-responses-to-all-human-viruses-using-infinity-bios)[![](https://cdn.sanity.io/images/vbcbvda6/production/42d91c8dac09c5fcb1af7609488897b8e4f36e5a-1200x750.webp)Brochure ### VirSIGHT InSIGHTs VirSIGHT at a glance: antibody reactome profiling across the known human virome (500+ species). Read more ](/resources/virsight-insights)[![](https://cdn.sanity.io/images/vbcbvda6/production/df6062d9a45026fd7525f846ef6c3f9c3f11fafb-1376x768.webp)Brochure ### Antibody Reactomics: The Responsive Layer of Immune Memory Where the antibody reactome fits in the multi-omic stack, how MIPSA deciphers it, and what the HuSIGHT, VirSIGHT, and En… Read more ](/resources/antibody-reactomics-overview)[![](https://cdn.sanity.io/images/vbcbvda6/production/bd98a023e7b99b56fcec6d9b14b0ba00a1435f06-1200x750.webp)Publication ### Viral Immunity in Immunoglobulin Products, Global Immunity Debt and Autoimmunity Lindahl et al., 2026: viral immunity in IgG products and global immunity debt. Read more ](/resources/viral-immunity-in-immunoglobulin-products-global-immunity-debt-and-autoimmunity)[![](https://cdn.sanity.io/images/vbcbvda6/production/bd98a023e7b99b56fcec6d9b14b0ba00a1435f06-1200x750.webp)Publication ### Applications of VirScan to broad serological profiling of bat reservoirs for emerging zoonoses Antibody reactome profiling of bat reservoirs identifies emerging zoonotic potential for biosurveillance. Read more ](/resources/applications-of-virscan-to-broad-serological-profiling-of-bat-reservoirs-for-eme)[![](https://cdn.sanity.io/images/vbcbvda6/production/bd98a023e7b99b56fcec6d9b14b0ba00a1435f06-1200x750.webp)Publication ### Comprehensive profiling of pre-infection antibodies identifies HIV targets associated with viremic control and viral load Frontiers Immunol 2023: pre-infection antibody reactome predicts post-infection HIV viral control. Read more ](/resources/comprehensive-profiling-of-pre-infection-antibodies-identifies-hiv-targets-assoc)[![](https://cdn.sanity.io/images/vbcbvda6/production/bd98a023e7b99b56fcec6d9b14b0ba00a1435f06-1200x750.webp)Publication ### Comprehensive profiling of antibody responses to the human anellome using programmable phage display Antibody reactome profiling of the human anellome: small DNA viruses persisting in healthy hosts. Read more ](/resources/comprehensive-profiling-of-antibody-responses-to-the-human-anellome-using-progra)[![](https://cdn.sanity.io/images/vbcbvda6/production/bd98a023e7b99b56fcec6d9b14b0ba00a1435f06-1200x750.webp)Publication ### Greater Breadth of Vaccine-Induced Immunity in Females than Males Is Mediated by Increased Antibody Diversity in Germinal Center B Cells Females develop broader vaccine-induced antibody repertoires, driven by germinal-center B-cell diversity. Read more ](/resources/greater-breadth-of-vaccine-induced-immunity-in-females-than-males-is-mediated-by)[![](https://cdn.sanity.io/images/vbcbvda6/production/bd98a023e7b99b56fcec6d9b14b0ba00a1435f06-1200x750.webp)Publication ### Longitudinal analysis reveals high prevalence of Epstein-Barr virus associated with multiple sclerosis Bjornevik et al., Science 2022: 10M serial serum samples link EBV to multiple sclerosis. Read more ](/resources/longitudinal-analysis-reveals-high-prevalence-of-epstein-barr-virus-associated-w)[![](https://cdn.sanity.io/images/vbcbvda6/production/bd98a023e7b99b56fcec6d9b14b0ba00a1435f06-1200x750.webp)Publication ### Analysis of antibody binding specificities in twin and SNP-genotyped cohorts reveals that antiviral antibody epitope selection is a heritable trait Antibody epitope selection is partially heritable, shown across twin and SNP-genotyped cohorts. Read more ](/resources/analysis-of-antibody-binding-specificities-in-twin-and-snp-genotyped-cohorts-rev)[![](https://cdn.sanity.io/images/vbcbvda6/production/bd98a023e7b99b56fcec6d9b14b0ba00a1435f06-1200x750.webp)Publication ### Deconvoluting virome-wide antibody epitope reactivity profiles Bioinformatic deconvolution of cross-reactive signals in virome-wide antibody reactome data. Read more ](/resources/deconvoluting-virome-wide-antibody-epitope-reactivity-profiles)[![](https://cdn.sanity.io/images/vbcbvda6/production/bd98a023e7b99b56fcec6d9b14b0ba00a1435f06-1200x750.webp)Publication ### SARS-CoV-2 antibody signatures robustly predict diverse antiviral functions relevant for convalescent plasma therapy SARS-CoV-2 antibody signatures predict convalescent plasma antiviral function. Read more ](/resources/sars-cov-2-antibody-signatures-robustly-predict-diverse-antiviral-functions-rele) Get in Touch [Contact Us](https://www.infinitybio.com/contact/)