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dc.contributor.authorCong, Yuehua
dc.contributor.authorPawlisz, Estera
dc.contributor.authorBryant, Penny
dc.contributor.authorBalan, Sibu
dc.contributor.authorLaurine, Emmanuelle
dc.contributor.authorTommasi, Rita
dc.contributor.authorSingh, Ruchi
dc.contributor.authorDubey, Sitara
dc.contributor.authorPeciak, Karolina
dc.contributor.authorBird, Matthew
dc.contributor.authorSivasankar, Amrita
dc.contributor.authorSwierkosz, Julia
dc.contributor.authorMuroni, Maurizio
dc.contributor.authorHeidelberger, Sibylle
dc.contributor.authorFarys, Monika
dc.contributor.authorKhayrzad, Farzad
dc.contributor.authorEdwards, Jeff
dc.contributor.authorBadescu, George
dc.contributor.authorHodgson, Ian
dc.contributor.authorHeise, Charles
dc.contributor.authorSomavarapu, Satyanarayana
dc.contributor.authorLiddell, John
dc.contributor.authorPowell, Keith
dc.contributor.authorZloh, Mire
dc.contributor.authorChoi, Ji-won
dc.contributor.authorGodwin, Antony
dc.contributor.authorBrocchini, Steve
dc.identifier.citationCong , Y , Pawlisz , E , Bryant , P , Balan , S , Laurine , E , Tommasi , R , Singh , R , Dubey , S , Peciak , K , Bird , M , Sivasankar , A , Swierkosz , J , Muroni , M , Heidelberger , S , Farys , M , Khayrzad , F , Edwards , J , Badescu , G , Hodgson , I , Heise , C , Somavarapu , S , Liddell , J , Powell , K , Zloh , M , Choi , J , Godwin , A & Brocchini , S 2012 , ' Site-specific PEGylation at histidine tags ' , Bioconjugate Chemistry , vol. 23 , no. 2 , pp. 248-63 .
dc.identifier.otherPURE: 1456221
dc.identifier.otherPURE UUID: e5adc71e-c934-4ab9-8c26-0d6582891819
dc.identifier.otherPubMed: 22243664
dc.identifier.otherScopus: 84863155417
dc.description.abstractThe efficacy of protein-based medicines can be compromised by their rapid clearance from the blood circulatory system. Achieving optimal pharmacokinetics is a key requirement for the successful development of safe protein-based medicines. Protein PEGylation is a clinically proven strategy to increase the circulation half-life of protein-based medicines. One limitation of PEGylation is that there are few strategies that achieve site-specific conjugation of PEG to the protein. Here, we describe the covalent conjugation of PEG site-specifically to a polyhistidine tag (His-tag) on a protein. His-tag site-specific PEGylation was achieved with a domain antibody (dAb) that had a 6-histidine His-tag on the C-terminus (dAb-His(6)) and interferon α-2a (IFN) that had an 8-histidine His-tag on the N-terminus (His(8)-IFN). The site of PEGylation at the His-tag for both dAb-His(6)-PEG and PEG-His(8)-IFN was confirmed by digestion, chromatographic, and mass-spectral studies. A methionine was also inserted directly after the N-terminal His-tag in IFN to give His(8)Met-IFN. Cyanogen bromide digestion studies of PEG-His(8)Met-IFN were also consistent with PEGylation at the His-tag. By using increased stoichiometries of the PEGylation reagent, it was possible to conjugate two separate PEG molecules to the His-tag of both the dAb and IFN proteins. Stability studies followed by in vitro evaluation confirmed that these PEGylated proteins retained their biological activity. In vivo PK studies showed that all of the His-tag PEGylated samples displayed extended circulation half-lives. Together, our results indicate that site-specific, covalent PEG conjugation at a His-tag can be achieved and biological activity maintained with therapeutically relevant proteins.en
dc.relation.ispartofBioconjugate Chemistry
dc.titleSite-specific PEGylation at histidine tagsen
dc.contributor.institutionSchool of Life and Medical Sciences
dc.contributor.institutionHealth & Human Sciences Research Institute
dc.contributor.institutionMedicinal and Analytical Chemistry
dc.contributor.institutionDepartment of Pharmacy
dc.description.statusPeer reviewed
dc.relation.schoolSchool of Life and Medical Sciences
rioxxterms.typeJournal Article/Review

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