earticle

영어

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.