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

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

doi:10.1038/s41598-020-71184-7

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

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

^*Corresponding author for this work

Pharmacology

Research output: Contribution to journal › Article › peer-review

13 Scopus citations

Abstract

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.

Original language	English (US)
Article number	14313
Journal	Scientific reports
Volume	10
Issue number	1
DOIs	https://doi.org/10.1038/s41598-020-71184-7
State	Published - Dec 1 2020

Funding

We are grateful to Ben Andrews and Randal Ketcham for insightful discussions. We thank Tsutomu Matsui and the staff at SSRL for assistance with SAXS data collection. We thank Dale Whittington and Scott Edgar for assistance with mass spectrometry data collection. This work was supported by the Global Health Vaccine Accelerator Program (GH-VAP #OPP1126258) by the Bill and Melinda Gates Foundation. SSRL is supported by the U.S. Department of Energy, Office of Science, Office of Basic Energy Sciences under Contract No. DE-AC02-76SF00515. The SSRL Structural Molecular Biology Program is supported by the DOE Office of Biological and Environmental Research, and by the National Institutes of Health, National Institute of General Medical Sciences (P41GM103393). GJR was funded as a Merck Fellow of the Life Sciences Research Foundation. BDW was funded by the Washington Research Foundation, Innovation Postdoctoral Fellowship.

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General

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This output contributes to the following UN Sustainable Development Goals (SDGs)

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title = "The influence of proline isomerization on potency and stability of anti-HIV antibody 10E8",

abstract = "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 YtransP conformation conformer can bind the HIV gp41 epitope, while the YcisP 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.",

author = "Miklos Guttman and Padte, {Neal N.} and Yaoxing Huang and Jian Yu and Rocklin, {Gabriel J.} and Weitzner, {Brian D.} and Michele Scian and Ho, {David D.} and Lee, {Kelly K.}",

note = "Funding Information: We are grateful to Ben Andrews and Randal Ketcham for insightful discussions. We thank Tsutomu Matsui and the staff at SSRL for assistance with SAXS data collection. We thank Dale Whittington and Scott Edgar for assistance with mass spectrometry data collection. This work was supported by the Global Health Vaccine Accelerator Program (GH-VAP #OPP1126258) by the Bill and Melinda Gates Foundation. SSRL is supported by the U.S. Department of Energy, Office of Science, Office of Basic Energy Sciences under Contract No. DE-AC02-76SF00515. The SSRL Structural Molecular Biology Program is supported by the DOE Office of Biological and Environmental Research, and by the National Institutes of Health, National Institute of General Medical Sciences (P41GM103393). GJR was funded as a Merck Fellow of the Life Sciences Research Foundation. BDW was funded by the Washington Research Foundation, Innovation Postdoctoral Fellowship. Publisher Copyright: {\textcopyright} 2020, The Author(s).",

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AU - Ho, David D.

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N2 - 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 YtransP conformation conformer can bind the HIV gp41 epitope, while the YcisP 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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The influence of proline isomerization on potency and stability of anti-HIV antibody 10E8

Abstract

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