Genetically encoded initiator for polymer growth from proteins

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United States of America Patent

PATENT NO 8816001
APP PUB NO 20130261259A1
SERIAL NO

13788710

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Abstract

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This invention pertains to methods for producing homogeneous recombinant proteins that contain polymer initiators at defined sites. The unnatural amino acid, 4-(2′-bromoisobutyramido)phenylalanine of formula 1, was designed and synthesized as a molecule comprising a functional group further comprising an initiator for an atom-transfer radical polymerization (‘ATRP”) that additionally would provide a stable linkage between the protein and growing polymer. We evolved a Methanococcus jannaschii (Mj) tyrosyl-tRNA synthetase/tRNACUA pair to genetically encode this unnatural amino acid in response to an amber codon. To demonstrate the utility of this functional amino acid, we produced Green Fluorescent Protein with the unnatural amino acid initiator of formula 1 site-specifically incorporated on its surface (GFP-1). Purified GFP-1 was then used as an initiator under standard ATRP conditions with oligo(ethylene oxide)monomethyl ether methacrylate, efficiently producing a polymer-GFP bioconjugate wherein the polymer is connected at a specifically selected site on GFP.

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Patent Owner(s)

Patent OwnerAddress
CARNEGIE MELLON UNIVERSITY5000 FORBES AVENUE PITTSBURGH PA 15213

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Inventor(s)

Inventor Name Address # of filed Patents Total Citations
Averick, Saadyah Pittsburgh, US 17 18
Matyjaszewski, Krzysztof Pittsburgh, US 92 3930
Mehl, Ryan A Corvallis, US 20 538

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