The influence of proline isomerization on potency and stability of anti-HIV antibody 10E8

Guttman, Miklos; Padte, Neal N.; Huang, Yaoxing; Yu, Jian; Rocklin, Gabriel J.; Weitzner, Brian D.; Scian, Michele; Ho, David D.; Lee, Kelly K.

doi:10.1038/s41598-020-71184-7

Title: The influence of proline isomerization on potency and stability of anti-HIV antibody 10E8

Journal Article · Mon Aug 31 00:00:00 EDT 2020 · Scientific Reports

DOI:https://doi.org/10.1038/s41598-020-71184-7· OSTI ID:1816235

Guttman, Miklos ^[1]; Padte, Neal N. ^[2]; Huang, Yaoxing ^[3]; Yu, Jian ^[3]; Rocklin, Gabriel J. ^[4]; Weitzner, Brian D. ^[5]; Scian, Michele ^[1]; Ho, David D. ^[3]; Lee, Kelly K. ^[1]

Univ. of Washington, Seattle, WA (United States). Dept. of Medicinal Chemistry
Columbia Univ., New York, NY (United States). Vagelos College of Physicians and Surgeons. Aaron Diamond AIDS Research Center
Columbia Univ., New York, NY (United States). Vagelos College of Physicians and Surgeons. Aaron Diamond AIDS Research Center
Northwestern Univ., Chicago, IL (United States). Dept. of Pharmacology
Univ. of Washington, Seattle, WA (United States). Dept. of Biochemistry; Univ. of Washington, Seattle, WA (United States). Inst. for Protein Design

Monoclonal antibody (mAb) 10E8 recognizes a highly conserved epitope on HIV and is capable of neutralizing> 95% of circulating viral isolates making it one of the most promising Abs against HIV. Solution instability and biochemical heterogeneity of 10E8 has hampered its development for clinical use. We identify the source of 10E8 heterogeneity being linked to cis/trans isomerization at two prolines within the YPP motif in the CRD3 loop that exists as two predominant conformers that interconvert on a slow timescale. The Y_transP conformation conformer can bind the HIV gp41 epitope, while the Y_cisP is not binding competent and shows a higher aggregation propensity. The high barrier of isomerization and propensity to adopt non-binding competent proline conformers provides novel insight into the slow binding kinetics, low potency, and poor solubility of 10E8. This study highlights how proline isomerization should be considered a critical quality attribute for biotherapeutics with paratopes containing potential cis proline amide bonds.

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Research Organization:: SLAC National Accelerator Lab., Menlo Park, CA (United States)

Sponsoring Organization:: USDOE Office of Science (SC), Basic Energy Sciences (BES). Scientific User Facilities Division; Bill and Melinda Gates Foundation; National Institutes of Health (NIH)

Grant/Contract Number:: AC02-76SF00515; P41GM103393; OPP1126258

OSTI ID:: 1816235

Journal Information:: Scientific Reports, Vol. 10, Issue 1; ISSN 2045-2322

Publisher:: Nature Publishing GroupCopyright Statement

Country of Publication:: United States

Language:: English

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Title: The influence of proline isomerization on potency and stability of anti-HIV antibody 10E8

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