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Title of Journal: Pharm Res

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Abbravation: Pharmaceutical Research

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Springer-Verlag

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10.1002/mop.22750

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1573-904X

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PEGylated Interferonα2b A Branched 40K Polyethyl

Authors: Jose Ramon Vivian Saez Reynier Baez Raymersy Aldana Eugenio Hardy
Publish Date: 2005/08/03
Volume: 22, Issue: 8, Pages: 1375-1387
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Abstract

We studied the conjugation of IFNα2b at different pH values 65 7 and 8 using the PEG240K reagent in either solution or solid state MonoPEGylated interferon was isolated by ionexchange chromatography and characterized using 1 sodium dodecyl sulfatepolyacrylamide gel electrophoresis 2 cation exchange highperformance liquid chromatography 3 bicinchoninic acid protein assay 4 enzymelinked immunosorbent assay 5 cellbased bioassays 6 thermal stability at 60°C 7 tryptic digestion and 8 pharmacokinetics in ratsPEGylation reaction gave 30–55 PEG240KIFNα2b 1–10 polyPEGylated interferon and 35–70 unmodified IFNα2b Compared to the polyPEGylated IFNα2b species the pure 96 monoPEGylated conjugate retained a significantly higher bioactivity IU/mg 17 × 104 ± 85 × 103 vs 28 × 106 ± 14 × 106 for antiviral and 19 × 104 ± 95 × 103 vs 31 × 106 ± 16 × 106 for antiproliferative activity Immunorecognition against IFN was reduced by the PEG240K moiety in the conjugate This monoPEGylated IFNα2b which migrated as a single band in gel electrophoresis was found to be a heterogeneous complex mixture of different positional isomers PEGylation markedly enhanced both the resistance to tryptic degradation and the thermal stability of IFNα2b The serum halflife of 40K PEG–IFN was 330fold longer while plasma residence time was increased 708 times compared to native IFN

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