Site-specific PEGylation of native disulfide bonds in therapeutic proteins.
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.
Shaunak
Sunil
S
Faculty of Medicine, Imperial College London, Hammersmith Hospital, Ducane Road, London W12 0NN, UK. s.shaunak@imperial.ac.uk
Godwin
Antony
A
Choi
Ji-Won
JW
Balan
Sibu
S
Pedone
Elisa
E
Vijayarangam
Damotharan
D
Heidelberger
Sibylle
S
Teo
Ian
I
Zloh
Mire
M
Brocchini
Steve
S
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
D000998:Antiviral Agents; D004220:Disulfides; D007140:Immunoglobulin Fab Fragments; D000077190:Interferon alpha-2; D016898:Interferon-alpha; D011994:Recombinant Proteins; D011092:Polyethylene Glycols; D003545:Cysteine