PRINT: A Protein Bioconjugation Method with Exquisite N-terminal Specificity

Surojit Sur, Yuan Qiao, Anja Fries, Robert N. O'Meally, Robert N Cole, Kenneth W Kinzler, Bert Vogelstein, Shibin Zhou

Research output: Contribution to journalArticle

Abstract

Chemical conjugation is commonly used to enhance the pharmacokinetics, biodistribution, and potency of protein therapeutics, but often leads to non-specific modification or loss of bioactivity. Here, we present a simple, versatile and widely applicable method that allows exquisite N-terminal specific modification of proteins. Combining reversible side-chain blocking and protease mediated cleavage of a commonly used HIS tag appended to a protein, we generate with high yield and purity exquisitely site specific and selective bio-conjugates of TNF-α by using amine reactive NHS ester chemistry. We confirm the N terminal selectivity and specificity using mass spectral analyses and show near complete retention of the biological activity of our model protein both in vitro and in vivo murine models. We believe that this methodology would be applicable to a variety of potentially therapeutic proteins and the specificity afforded by this technique would allow for rapid generation of novel biologics.

Original languageEnglish (US)
Article number18363
JournalScientific Reports
Volume5
DOIs
StatePublished - Dec 17 2015

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Proteins
Biological Products
Amines
Esters
Peptide Hydrolases
Pharmacokinetics
Therapeutics
In Vitro Techniques

ASJC Scopus subject areas

  • General

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PRINT : A Protein Bioconjugation Method with Exquisite N-terminal Specificity. / Sur, Surojit; Qiao, Yuan; Fries, Anja; O'Meally, Robert N.; Cole, Robert N; Kinzler, Kenneth W; Vogelstein, Bert; Zhou, Shibin.

In: Scientific Reports, Vol. 5, 18363, 17.12.2015.

Research output: Contribution to journalArticle

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