Abstract

The broadly neutralizing antibody (bNAb) CAP256-VRC26.25 has exceptional potency against HIV-1 and has been considered for clinical use. During the characterization and production of this bNAb, we observed several unusual features. First, the antibody appeared to adhere to pipette tips, requiring tips to be changed during serial dilution to accurately measure potency. Second, during production scale-up, proteolytic cleavage was discovered to target an extended heavy chain loop, which was attributed to a protease in spent medium from 2-week culture. To enable large scale production, we altered the site of cleavage via a single amino acid change, K100mA. The resultant antibody retained potency and breadth while avoiding protease cleavage. We also added the half-life extending mutation LS, which improved the in vivo persistence in animal models, but did not impact neutralization activity; we observed the same preservation of neutralization for bNAbs VRC01, N6, and PGDM1400 with LS on a 208-virus panel. The final engineered antibody, CAP256V2LS, retained the extraordinary neutralization potency of the parental antibody, had a favorable pharmacokinetic profile in animal models, and was negative in in vitro assessment of autoreactivity. CAP256V2LS has the requisite potency, developability and suitability for scale-up, allowing its advancement as a clinical candidate.

Details

Title
Engineering of HIV-1 neutralizing antibody CAP256V2LS for manufacturability and improved half life
Author
Zhang, Baoshan 1 ; Gollapudi, Deepika 1 ; Gorman, Jason 1 ; O’Dell, Sijy 1 ; Damron, Leland F. 1 ; McKee, Krisha 1 ; Asokan, Mangaiarkarasi 1 ; Yang, Eun Sung 1 ; Pegu, Amarendra 1 ; Lin, Bob C. 1 ; Chao, Cara W. 1 ; Chen, Xuejun 1 ; Gama, Lucio 1 ; Ivleva, Vera B. 1 ; Law, William H. 1 ; Liu, Cuiping 1 ; Louder, Mark K. 1 ; Schmidt, Stephen D. 1 ; Shen, Chen-Hsiang 1 ; Shi, Wei 1 ; Stein, Judith A. 1 ; Seaman, Michael S. 2 ; McDermott, Adrian B. 1 ; Carlton, Kevin 1 ; Mascola, John R. 1 ; Kwong, Peter D. 1 ; Lei, Q. Paula 1 ; Doria-Rose, Nicole A. 1 

 National Institutes of Health, Vaccine Research Center, National Institute of Allergy and Infectious Diseases, Bethesda, USA (GRID:grid.94365.3d) (ISNI:0000 0001 2297 5165) 
 Beth Israel Deaconess Medical Center, Boston, USA (GRID:grid.239395.7) (ISNI:0000 0000 9011 8547); Harvard Medical School, Boston, USA (GRID:grid.38142.3c) (ISNI:000000041936754X) 
Publication year
2022
Publication date
2022
Publisher
Nature Publishing Group
e-ISSN
20452322
Source type
Scholarly Journal
Language of publication
English
ProQuest document ID
2728335443
Copyright
© This is a U.S. Government work and not under copyright protection in the US; foreign copyright protection may apply 2022. 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.