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

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 model

SUBMITTER: Sur S 

PROVIDER: S-EPMC4683619 | biostudies-literature | 2015 Dec

REPOSITORIES: biostudies-literature