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.

Details

Title
The influence of proline isomerization on potency and stability of anti-HIV antibody 10E8
Author
Guttman Miklos 1 ; Padte, Neal N 2 ; Huang Yaoxing 2 ; Yu, Jian 2 ; Rocklin, Gabriel J 3 ; Weitzner, Brian D 4 ; Scian Michele 1 ; Ho, David D 2 ; Lee, Kelly K 1 

 University of Washington, Department of Medicinal Chemistry, Seattle, USA (GRID:grid.34477.33) (ISNI:0000000122986657) 
 Columbia University Vagelos College of Physicians and Surgeons, Aaron Diamond AIDS Research Center, New York, USA (GRID:grid.21729.3f) (ISNI:0000000419368729) 
 Northwestern University, Department of Pharmacology, Chicago, USA (GRID:grid.16753.36) (ISNI:0000 0001 2299 3507) 
 University of Washington, Department of Biochemistry, Seattle, USA (GRID:grid.34477.33) (ISNI:0000000122986657); University of Washington, Institute for Protein Design, Seattle, USA (GRID:grid.34477.33) (ISNI:0000000122986657) 
Publication year
2020
Publication date
2020
Publisher
Nature Publishing Group
e-ISSN
20452322
Source type
Scholarly Journal
Language of publication
English
ProQuest document ID
1886292134
Copyright
© The Author(s) 2020. This work is published under http://creativecommons.org/licenses/by/4.0/ (the “License”). Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.