Staff Publications

Staff Publications

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    'Staff publications' is the digital repository of Wageningen University & Research

    'Staff publications' contains references to publications authored by Wageningen University staff from 1976 onward.

    Publications authored by the staff of the Research Institutes are available from 1995 onwards.

    Full text documents are added when available. The database is updated daily and currently holds about 240,000 items, of which 72,000 in open access.

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Record number 551774
Title Inducible, selective labeling of proteins via enzymatic oxidation of tyrosine
Author(s) Bruins, Jorick J.; Wouw, Criss van de; Keijzer, Jordi F.; Albada, Bauke; Delft, Floris L. van
Source In: Methods in Molecular Biology Hertfordshire : Springer (Methods in Molecular Biology ) - ISBN 9781493995455 - p. 357 - 368.
DOI https://doi.org/10.1007/978-1-4939-9546-2_18
Department(s) Organic Chemistry
VLAG
Publication type Peer reviewed book chapter
Publication year 2019
Keyword(s) Antibody–drug conjugates (ADC) - Bioconjugate chemistry - Bioorthogonal click - Protein functionalization - SPOCQ - Tyrosine modification
Abstract

Proteins can be labeled site-specifically and in inducible fashion by exposing a small peptide tag (G4Y) on any of its termini and activating the newly exposed tyrosine residue with the enzyme mushroom tyrosinase. The enzyme generates a quinone by oxidizing the tyrosine, which in turn can perform strain-promoted oxidation-controlled ortho-quinone cycloaddition (SPOCQ) with strained alkynes and alkenes, generating a stable conjugation product. Here, we describe a protocol to perform SPOCQ reaction on proteins, along with notes to optimize yield and reaction rates. Conjugation efficiencies of over 95% to antibodies have been reported using this protocol.

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