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.
journal_name
Nat Biotechnoljournal_title
Nature biotechnologyauthors
Shen BQ,Xu K,Liu L,Raab H,Bhakta S,Kenrick M,Parsons-Reponte KL,Tien J,Yu SF,Mai E,Li D,Tibbitts J,Baudys J,Saad OM,Scales SJ,McDonald PJ,Hass PE,Eigenbrot C,Nguyen T,Solis WA,Fuji RN,Flagella KM,Patel D,Spedoi
10.1038/nbt.2108subject
Has Abstractpub_date
2012-01-22 00:00:00pages
184-9issue
2eissn
1087-0156issn
1546-1696pii
nbt.2108journal_volume
30pub_type
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