CAJ | 학술논문

目的采用凝胶过滤层析法分离全蝎酶解液的抗凝活性部位,并检测活性部分的分子量分布范围.方法采用紫外检测法、考马斯亮蓝法检测,合并凝胶层析蛋白流分;以APTT凝血时间为指标,检验各组分体外抗凝活性;通过MALDI-TOF-MASS法检测分离部分的分子量分布范围.结果全蝎酶解液通过凝胶过滤层析分得的2组分抗凝效果显著,分子量分布范围850～5 300.结论体外抗凝试验表明:凝胶过滤层析法纯化分离全蝎胃蛋白酶酶解液方法可行,MALDI-TOF-MASS表明活性组分为小分子肽类.
목적 채용응효과려층석법분리전갈매해액적항응활성부위,병검측활성부분적분자량분포범위.방법 채용자외검측법、고마사량람법검측,합병응효층석단백류분;이APTT응혈시간위지표,검험각조분체외항응활성;통과MALDI-TOF-MASS법검측분리부분적분자량분포범위.결과 전갈매해액통과응효과려층석분득적2조분항응효과현저,분자량분포범위850～5 300.결론 체외항응시험표명:응효과려층석법순화분리전갈위단백매매해액방법 가행,MALDI-TOF-MASS표명활성조분위소분자태류.
Objective To separate the fractions with anticoagulant activity from the pepsin hydrolytic liquid of Quanxie (Scorpio), and detect the distribution of molecular weight in the active fractions by applying gel filtration chromatography.Methods The methods of UV-detection and Coomassie brilliant blue G250 were used to detect and combine protein fractions of gel chromatography. The anticoagulant activities of different fractions were detected taken activated partial thromboplastin time (APTT) as index. The distribution of molecular weight of the separated active fractions was determined by using MALDI-TOF-MS.Results There two fractions, separated from the pepsin hydrolytic liquid of Quanxie by gel filtration chromatography, with significant anticoagulant activities. The molecular weight distributed from 850 to 5 300.Conclusion The result of anticoagulant test in vitro indicates that gel filtration chromatography is feasible for purifying and separating the pepsin hydrolytic liquid of Quanxie. The outcomes of MALDI-TOF-MS show that the active fraction is micromolecular peptide.