Ou, Weijia, Bursulaya, Badry, Didonato, Michael, Herath, Ananda, Jin, Yunho, Hao, Xueshi, Loren, Jon, Brock, Ansgar, Uno, Tetsuo, Geierstanger, Bernhard, Cellitti, Susan and Mortenson, Grace (2018) Tuning a Protein-Labeling Reaction to Achieve Highly Site Selective Lysine Conjugation. ChemBioChem, 19 (8). pp. 799-804. ISSN 14397633

Abstract

Activated esters are widely used to label proteins at lysine side chains and N termini. These reagents are useful for labeling virtually any protein, but robust reactivity toward primary amines generally precludes site-selective modification. In a unique case, fluorophenyl esters are shown to preferentially label human kappa antibodies at a single lysine (Lys188) within the light-chain constant domain. Neighboring residues His189 and Asp151 contribute to the accelerated rate of labeling at Lys188 relative to the ≈40 other lysine sites. Enriched Lys188 labeling can be enhanced from 50–70 % to >95 % by any of these approaches: lowering reaction temperature, applying flow chemistry, or mutagenesis of specific residues in the surrounding protein environment. Our results demonstrated that activated esters with fluoro-substituted aromatic leaving groups, including a fluoronaphthyl ester, can be generally useful reagents for site-selective lysine labeling of antibodies and other immunoglobulin-type proteins.

Item Type:	Article
Keywords:	antibodies bioconjugation chemical tools flow chemistry lysine
Date Deposited:	19 Jun 2018 00:45
Last Modified:	25 Jan 2019 00:45
URI:	https://oak.novartis.com/id/eprint/34545