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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-\u03b1 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 <jats:italic>in vitro<\/jats:italic> and <jats:italic>in vivo<\/jats:italic> 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.<\/jats:p>","DOI":"10.1038\/srep18363","type":"journal-article","created":{"date-parts":[[2015,12,18]],"date-time":"2015-12-18T12:27:27Z","timestamp":1450441647000},"update-policy":"http:\/\/dx.doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["PRINT: A Protein Bioconjugation Method with Exquisite N-terminal Specificity"],"prefix":"10.1038","volume":"5","author":[{"given":"Surojit","family":"Sur","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yuan","family":"Qiao","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Anja","family":"Fries","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Robert N.","family":"O\u2019Meally","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Robert N.","family":"Cole","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kenneth W.","family":"Kinzler","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Bert","family":"Vogelstein","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Shibin","family":"Zhou","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"297","published-online":{"date-parts":[[2015,12,17]]},"reference":[{"key":"BFsrep18363_CR1","doi-asserted-by":"publisher","first-page":"507","DOI":"10.1038\/nrd2039","volume":"5","author":"C Krejsa","year":"2006","unstructured":"Krejsa, C., Rogge, M. & Sadee, W. 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