Author:
Zhang Baoshan,Gollapudi Deepika,Gorman Jason,O’Dell Sijy,Damron Leland F.,McKee Krisha,Asokan Mangaiarkarasi,Yang Eun Sung,Pegu Amarendra,Lin Bob C.,Chao Cara W.,Chen Xuejun,Gama Lucio,Ivleva Vera B.,Law William H.,Liu Cuiping,Louder Mark K.,Schmidt Stephen D.,Shen Chen-Hsiang,Shi Wei,Stein Judith A.,Seaman Michael S.,McDermott Adrian B.,Carlton Kevin,Mascola John R.,Kwong Peter D.,Lei Q. Paula,Doria-Rose Nicole A.
Abstract
AbstractThe 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.
Funder
Division of Intramural Research, National Institute of Allergy and Infectious Diseases
National Institutes of Health
Publisher
Springer Science and Business Media LLC