| Literature DB >> 22267010 |
Ben-Quan Shen1, Keyang Xu, Luna Liu, Helga Raab, Sunil Bhakta, Margaret Kenrick, Kathryn L Parsons-Reponte, Janet Tien, Shang-Fan Yu, Elaine Mai, Dongwei Li, Jay Tibbitts, Jakub Baudys, Ola M Saad, Suzie J Scales, Paul J McDonald, Philip E Hass, Charles Eigenbrot, Trung Nguyen, Willy A Solis, Reina N Fuji, Kelly M Flagella, Darshana Patel, Susan D Spencer, Leslie A Khawli, Allen Ebens, Wai Lee Wong, Richard Vandlen, Surinder Kaur, Mark X Sliwkowski, Richard H Scheller, Paul Polakis, Jagath R Junutula.
Abstract
The reactive thiol in cysteine is used for coupling maleimide linkers in the generation of antibody conjugates. To assess the impact of the conjugation site, we engineered cysteines into a therapeutic HER2/neu antibody at three sites differing in solvent accessibility and local charge. The highly solvent-accessible site rapidly lost conjugated thiol-reactive linkers in plasma owing to maleimide exchange with reactive thiols in albumin, free cysteine or glutathione. In contrast, a partially accessible site with a positively charged environment promoted hydrolysis of the succinimide ring in the linker, thereby preventing this exchange reaction. The site with partial solvent-accessibility and neutral charge displayed both properties. In a mouse mammary tumor model, the stability and therapeutic activity of the antibody conjugate were affected positively by succinimide ring hydrolysis and negatively by maleimide exchange with thiol-reactive constituents in plasma. Thus, the chemical and structural dynamics of the conjugation site can influence antibody conjugate performance by modulating the stability of the antibody-linker interface.Entities:
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Year: 2012 PMID： 22267010 DOI： 10.1038/nbt.2108
Source DB: PubMed Journal: Nat Biotechnol ISSN： 1087-0156 Impact factor: 54.908