N-terminal site-specific mono-PEGylation of epidermal growth factor.

Abstract:

PURPOSE:N-terminal site-specific mono-PEGylation of recombinant human epidermal growth factor (EGF) was accomplished using polyethyleneglycol (PEG) derivatives (Mw = 2000 and 5000) through a reactive terminal aldehyde group. METHODS:The site-specific PEG conjugation was conducted ata slightly acidic pH condition (pH 5.5). The mono-PEGylation was targeted to an alpha-amine group at the N-terminal end of EGF to minimize reduction of biologic activity. Tryptic digestion mapping and MALDI-TOF MS techniques were applied to show the occurrence of mono-PEGylation at the N-terminus of EGF. RESULTS:The site-specific mono-PEGylated EGF, when compared with native EGF, fully retained its in vitro biologic activities such a cell proliferation and intracellular signal transduction. This revealed that although a synthetic polymer of a PEG was covalently conjugated to EGF, the internalized complex of PEGylated EGF-receptor within cells did not hamper the intracellular signal transduction events. The PEGylated EGF also exhibited a prolonged circulation in blood stream in vivo and markedly enhanced physical stability whe incubated with tissue homogenate. CONCLUSION:N-terminally mono-PEGylated EGF shows increased physical stability while retaining its biologic activity.

journal_name

Pharm Res

journal_title

Pharmaceutical research

authors

Lee H,Jang IH,Ryu SH,Park TG

doi

10.1023/a:1023402123119

subject

Has Abstract

pub_date

2003-05-01 00:00:00

pages

818-25

issue

5

eissn

0724-8741

issn

1573-904X

journal_volume

20

pub_type

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