biostudies-literatureSajadi MMBLRD VANational Institute of Allergy and Infectious Diseases Division of Intramural ResearchNIAID NIH HHSUniversity of MarylandBiological and Environmental ResearchOffice of ScienceNational Institutes of HealthNational Institute of General Medical SciencesUS Department of Veterans AffairsNational Institute of Allergy and Infectious DiseasesU.S. Department of EnergyNational Institute of Standards and TechnologyBasic Energy SciencesGates FoundationNIGMS NIH HHS1150-1164.e8https://www.ebi.ac.uk/biostudies/studies/S-EPMC12227296biostudies-literatureUnknown33(7)Anti-HIV envelope broadly neutralizing antibodies (bnAbs) are alternatives to conventional antiretrovirals with the potential to prevent and treat infection, reduce latent reservoirs, and/or mediate a functional cure. Clinical trials with "first-generation" bnAbs used alone or in combination show promising antiviral effects but also highlight that additional engineering of "enhanced" antibodies will be required for optimal clinical utility, while preserving or enhancing Current Good Manufacturing Practices (cGMP) manufacturing capability. Here, we report the engineering of an anti-CD4-binding site (CD4bs) bnAb, N49P9.3. Through a series of rational modifications, we produced a variant, N49P9.6-FR-LS, that demonstrates enhanced potency, superior antiviral activity in combination with other Structure (London, England : 1993)A comprehensive engineering strategy improves potency and manufacturability of a near pan-neutralizing antibody against HIV.PMC1222729670NANB21H105INV-036842R01 AI155150I01 BX004525DE-AC02-76SF00515R35 GM144083IBX004525P01 AI1622421R01AI155150-01A1INV-005284R01 AI174908R01 AI147870R01AI147870Pierce BGHabibzadeh PKarimi MJardine JGSaadat SOuyang XAtkinson BMielke DDunn YLiu QTehrani ZRWagh KPozharski ETolbert WDZareidoodeji RMontefiori DKedzior MFerrer DSeaman MSSajadi MMSprague MChi WDeVico AAslanabadi AAbbasi APazgier MHeredia AYin RLewis GKFerrari GKorber BChen HNguyen DNGuest JDSiska CJumpa TMayer BTKetchem RRCalvillo SAsakawa AClark RDeming DFoulke SLusso PSato AHHojabri MfalseA comprehensive engineering strategy improves potency and manufacturability of a near pan-neutralizing antibody against HIV.Anti-HIV envelope broadly neutralizing antibodies (bnAbs) are alternatives to conventional antiretrovirals with the potential to prevent and treat infection, reduce latent reservoirs, and/or mediate a functional cure. Clinical trials with "first-generation" bnAbs used alone or in combination show promising antiviral effects but also highlight that additional engineering of "enhanced" antibodies will be required for optimal clinical utility, while preserving or enhancing Current Good Manufacturing Practices (cGMP) manufacturing capability. Here, we report the engineering of an anti-CD4-binding site (CD4bs) bnAb, N49P9.3. Through a series of rational modifications, we produced a variant, N49P9.6-FR-LS, that demonstrates enhanced potency, superior antiviral activity in combination with other 2025-01-01T00:00:00Z2025 Jul2026-06-02T08:53:32.385Z2026-04-16T03:12:38.28ZS-EPMC122272964037376610.1016/j.str.2025.04.016