TY - JOUR
T1 - Engineering of HIV-1 neutralizing antibody CAP256V2LS for manufacturability and improved half life
AU - Zhang, Baoshan
AU - Gollapudi, Deepika
AU - Gorman, Jason
AU - O’Dell, Sijy
AU - Damron, Leland F.
AU - McKee, Krisha
AU - Asokan, Mangaiarkarasi
AU - Yang, Eun Sung
AU - Pegu, Amarendra
AU - Lin, Bob C.
AU - Chao, Cara W.
AU - Chen, Xuejun
AU - Gama, Lucio
AU - Ivleva, Vera B.
AU - Law, William H.
AU - Liu, Cuiping
AU - Louder, Mark K.
AU - Schmidt, Stephen D.
AU - Shen, Chen Hsiang
AU - Shi, Wei
AU - Stein, Judith A.
AU - Seaman, Michael S.
AU - McDermott, Adrian B.
AU - Carlton, Kevin
AU - Mascola, John R.
AU - Kwong, Peter D.
AU - Lei, Q. Paula
AU - Doria-Rose, Nicole A.
N1 - Funding Information:
The authors thank Cindy Cai for technical assistance, and Brenda Hartman for assistance with figures and manuscript preparation. They thank J. Baalwa, D. Ellenberger, F. Gao, B. Hahn, K. Hong, J. Kim, F. McCutchan, D. Montefiori, L. Morris, J. Overbaugh, E. Sanders-Buell, G. Shaw, R. Swanstrom, M. Thomson, S. Tovanabutra, C. Williamson and L. Zhang for contributing the HIV-1 Envelope plasmids used in our neutralization panel. The master cell bank for production of PGT121.414.LS was provided by Beth Israel Deaconess Medical Center. Funding was provided by the Intramural Research Programs of the Vaccine Research Center, National Institute of Allergy and Infectious Diseases, National Institutes of Health, USA.
Publisher Copyright:
© 2022, This is a U.S. Government work and not under copyright protection in the US; foreign copyright protection may apply.
PY - 2022/12
Y1 - 2022/12
N2 - 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.
AB - 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.
UR - https://www.scopus.com/pages/publications/85140654163
UR - https://www.scopus.com/pages/publications/85140654163#tab=citedBy
U2 - 10.1038/s41598-022-22435-2
DO - 10.1038/s41598-022-22435-2
M3 - Article
C2 - 36284200
AN - SCOPUS:85140654163
SN - 2045-2322
VL - 12
JO - Scientific reports
JF - Scientific reports
IS - 1
M1 - 17876
ER -