Site-Specific Antibody Functionalization Using Tetrazine–Styrene Cycloaddition
Name
nihms966512.pdf
Description
Accepted version
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642.31 KB
Format
Adobe PDF
Checksum (MD5)
dbe6c22ceb2eb2f2898211ac9e23228a
Author(s)
Mix, Kalie
Raines, Ronald T
Date Issued
May 2018
Journal
Bioconjugate chemistry
Publisher
American Chemical Society (ACS)
Citation
Umlauf, Benjamin J. et al. “Site-Specific Antibody Functionalization Using Tetrazine–Styrene Cycloaddition.” Bioconjugate chemistry 29 (2018): 1605-1613 © 2018 The Author(s)
Version
Author's final manuscript
Abstract
Biologics, such as antibody-drug conjugates, are becoming mainstream therapeutics. Consequently, methods to functionalize biologics without disrupting their native properties are essential for identifying, characterizing, and translating candidate biologics from the bench to clinical practice. Here, we present a method for site-specific, carboxy-terminal modification of single-chain antibody fragments (scFvs). ScFvs displayed on the surface of yeast were isolated and functionalized by combining intein-mediated expressed protein ligation (EPL) with inverse electron-demand Diels-Alder (IEDDA) cycloaddition using a styrene-tetrazine pair. The high thiol concentration required to trigger EPL can hinder the subsequent chemoselective ligation reactions; therefore, the EPL reaction was used to append styrene to the scFv, limiting tetrazine exposure to damaging thiols. Subsequently, the styrene-functionalized scFv was reacted with tetrazine-conjugated compounds in an IEDDA cycloaddition to generate functionalized scFvs that retain their native binding activity. Rapid functionalization of yeast surface-derived scFv in a site-directed manner could find utility in many downstream laboratory and preclinical applications.
MIT Department
Massachusetts Institute of Technology. Department of Chemistry
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Article is made available in accordance with the publisher's policy and may be subject to US copyright law. Please refer to the publisher's site for terms of use.
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DOI of Published Version
https://doi.org/10.1021/ACS.BIOCONJCHEM.8B00114
