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재조합 소성장호르몬의 전체 아미노산 서열분석을 위한 2 차원 고성능 액체 크로마토그래피법에 관한 연구
김정호,김훈주,김준 ( Jungho Kim,Hoon Joo Kim,Joon Kim ) 생화학분자생물학회 1992 BMB Reports Vol.25 No.3
In order to separate and purify tryptic digested recombinant bST (rbST), 2-dimensional HPLC which involves a combination of strong cation-exchange chromatography and reverse-phase chromatography was applied. A single dimension RP-HPLC resulted in several heterogeneous peaks. However, all fractions purified by two-dimensional chromatogrphy resulted in homogeneous peptides and unambiguous sequence data. With a view of separating perfectly, tryptic digest was first fractionated by strong cation-exchange chromatogrphy. The nine fractions were obtained from this chromatogrphy and each of these fractions was applied to RP-HPLC. Fractions yielded from RP-HPLC were submitted for direct sequence analysis and clear sequence data were obtained. In this study, the location of disulfide bonds, Cys53-Cys164 and Cys181-Cys189, also were confirmed in rbST.