Conjugation Site Modulates the in Vivo Stability and Therapeutic Activity of Antibody-drug Conjugates

Overview

Journal Nat Biotechnol

Specialty Biotechnology

Date 2012 Jan 24

PMID 22267010

Citations 305

Authors

Ben-Quan Shen

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

Affiliations

Soon will be listed here.

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.

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