Engineering humanized antibody framework sequences for optimal site-specific conjugation of cytotoxins.

Autor:	Spidel JL; a Morphotek Inc. , Exton , PA , USA., Albone EF; a Morphotek Inc. , Exton , PA , USA., Cheng X; a Morphotek Inc. , Exton , PA , USA., Vaessen B; a Morphotek Inc. , Exton , PA , USA., Jacob S; a Morphotek Inc. , Exton , PA , USA., Milinichik AZ; a Morphotek Inc. , Exton , PA , USA., Verdi A; a Morphotek Inc. , Exton , PA , USA., Kline JB; a Morphotek Inc. , Exton , PA , USA., Grasso L; a Morphotek Inc. , Exton , PA , USA.
Jazyk:	angličtina
Zdroj:	MAbs [MAbs] 2017 Aug/Sep; Vol. 9 (6), pp. 907-915. Date of Electronic Publication: 2017 May 25.
DOI:	10.1080/19420862.2017.1330734
Abstrakt:	The prevailing techniques used to generate antibody-drug conjugates (ADCs) involve random conjugation of the linker-drug to multiple lysines or cysteines in the antibody. Engineering natural and non-natural amino acids into an antibody has been demonstrated to be an effective strategy to produce homogeneous ADC products with defined drug-to-antibody ratios. We recently reported an efficient residue-specific conjugation technology (RESPECT) where thiol-reactive payloads can be efficiently conjugated to a native unpaired cysteine in position 80 (C80) of rabbit light chains. Deimmunizing the rabbit variable domains through humanization is necessary to reduce the risk of anti-drug antibody responses in patients. However, we found that first-generation humanized RESPECT ADCs showed high levels of aggregation and low conjugation efficiency. We correlated these negative properties to the phenylalanine at position 83 present in most human variable kappa frameworks. When position 83 was substituted with selected amino acids, conjugation was restored and aggregation was reduced to levels similar to the chimeric ADC. This engineering strategy allows for development of second-generation humanized RESPECT ADCs with desirable biopharmaceutical properties.
Databáze:	MEDLINE
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