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N-Terminal Site-Specific Mono-PEGylation of Epidermal Growth Factor

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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 at a slightly acidic pH condition (pH 5.5). The mono-PEGylation was targeted to an α-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 as 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 when incubated with tissue homogenate.

Conclusion. N-terminally mono-PEGylated EGF shows increased physical stability while retaining its biologic activity.

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Authors and Affiliations

  1. Department of Biologic Sciences, Korea Advanced Institute of Science and Technology Daejeon, 305-701, Korea

    Haeshin Lee & Tae Gwan Park

  2. Division of Molecular and, Life Science Pohang University of Science and Technology Pohang, 790-784, Korea

    Il Ho Jang & Sung Ho Ryu

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  1. Haeshin Lee
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Lee, H., Jang, I.H., Ryu, S.H. et al. N-Terminal Site-Specific Mono-PEGylation of Epidermal Growth Factor. Pharm Res 20, 818–825 (2003). https://doi.org/10.1023/A:1023402123119

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