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Conjugation site modulates the in vivo stability and therapeutic activity of antibody-drug conjugates.

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

Nat Biotechnol 2012 Jan 22;30(2):184-9. Epub 2012 Jan 22.

Genentech Inc., 1 DNA Way, S. San Francisco, California, USA.

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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http://dx.doi.org/10.1038/nbt.2108DOI Listing
January 2012

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