title: Site-specific PEGylation of native disulfide bonds in therapeutic proteins.
abstract: Native disulfide bonds in therapeutic proteins are crucial for tertiary structure and biological activity and are therefore considered unsuitable for chemical modification. We show that native disulfides in human interferon alpha-2b and in a fragment of an antibody to CD4(+) can be modified by site-specific bisalkylation of the two cysteine sulfur atoms to form a three-carbon PEGylated bridge. The yield of PEGylated protein is high, and tertiary structure and biological activity are retained.
authors.affiliation: Faculty of Medicine, Imperial College London, Hammersmith Hospital, Ducane Road, London W12 0NN, UK. s.shaunak@imperial.ac.uk
authors.forename: Sunil
authors.initials: S
authors.lastname: Shaunak
authors.forename: Antony
authors.initials: A
authors.lastname: Godwin
authors.forename: Ji-Won
authors.initials: JW
authors.lastname: Choi
authors.forename: Sibu
authors.initials: S
authors.lastname: Balan
authors.forename: Elisa
authors.initials: E
authors.lastname: Pedone
authors.forename: Damotharan
authors.initials: D
authors.lastname: Vijayarangam
authors.forename: Sibylle
authors.initials: S
authors.lastname: Heidelberger
authors.forename: Ian
authors.initials: I
authors.lastname: Teo
authors.forename: Mire
authors.initials: M
authors.lastname: Zloh
authors.forename: Steve
authors.initials: S
authors.lastname: Brocchini
mesh_terms: D000478:Alkylation; D000998:Antiviral Agents / Q000737:chemistry* / Q000378:metabolism / Q000494:pharmacology; D001665:Binding Sites; D002460:Cell Line; D003545:Cysteine / Q000737:chemistry / Q000378:metabolism; D004220:Disulfides / Q000737:chemistry* / Q000378:metabolism; D015497:HIV-1 / Q000187:drug effects; D006801:Humans; D007140:Immunoglobulin Fab Fragments / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D000077190:Interferon alpha-2; D016898:Interferon-alpha / Q000737:chemistry* / Q000378:metabolism / Q000494:pharmacology; D015394:Molecular Structure; D011092:Polyethylene Glycols / Q000737:chemistry* / Q000378:metabolism; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins; D013329:Structure-Activity Relationship
chemical_list: D000998:Antiviral Agents; D004220:Disulfides; D007140:Immunoglobulin Fab Fragments; D000077190:Interferon alpha-2; D016898:Interferon-alpha; D011994:Recombinant Proteins; D011092:Polyethylene Glycols; D003545:Cysteine