PEGylation of biopharmaceutical proteins is important to increase in serum circulation stabilityand to minimize antigenicity. The most preferable method of PEGylation is a site-orterminal-specific, mono-EGylation. We propose the terminal-specific mono-PEGylationmethod using intein-mediated fusion protein technology. By exploiting the affinity taggingdomain (chitin binding domain) of a fusion protein, we were able to immobilize the C-terminusof the fusion protein to chitin matrix, exposing the N-terminus for solid-phase PEGylation.The distinct advantage of inteins is self-splicing ability by simple changes in pH and/ortemperature without the need for cleavage proteins or reagents. By this way, we couldpresent an integrated process for expression-refolding-PEGylation-purification. Rh-EGF(recombinant human epidermal growth factor) was used as a model protein. The PEGylationsite was determined by mass spectrometry, and the bioactivity of the modified rhEGF wascompared with the native and randomly PEGylated EGF using NRK cell culture assay.
키워드
inteinterminal-specificmono-PEGylationrhEGF
저자
Jung Hye KANG [ Bioprocessing Research Laboratory, Dept. of Chemical Engineering, Hanyang University, Ansan, 426-791. ]
HyunKyoung BYUN [ Bioprocessing Research Laboratory, Dept. of Bionano technology, Hanyang University, Ansan, 426-791. ]
Eun Kyu LEE [ College of Bionanotechnology, Kyung Won University, Sungnam, 461-701. ]
한국생물공학회 [The Korean Society for Biotechnology and Bioengineering]
설립연도
1984
분야
공학>생물공학
소개
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