脱细胞猪小肠黏膜下层基质中纤维连接蛋白定量检测方法研究  

Quantitative detection of fibronectin in porcine small intestinal submucosa extracellular matrix

作　　者：陈毅 夏磊磊 张扬 高建萍[3] 李赛娜 雷雄心 李勇超[2] 张贵锋[3] 赵博 CHEN Yi;XIA Lei-lei;ZHANG Yang;GAO Jian-ping;LI Sai-na;LEI Xiong-xin;LI Yong-chao;ZHANG Gui-feng;ZHAO Bo(Beijing Biosis Healing Biological Technology Co. , Ltd. , Beijing 100026;State Key Laboratory of Biochemical Engineering, Institute of Process Engineering, CAS, Beijing 100190;School of Life Science and Technology, Henan Institute of Science and Technology, Xinxiang 453003, China)

机构地区：[1]北京博辉瑞进生物科技有限公司,北京100026 [2]河南科技学院生命科技学院,新乡453003 [3]中国科学院过程工程研究所生化工程国家重点实验室,北京100190

出　　处：《生物学杂志》2018年第2期97-100,共4页Journal of Biology

基　　金：北京市科技新星计划(Z181100006218013;XX2018008);国家863计划新材料领域专项(2015AA033602);科技部中小企业创新基金(Z14010101281);广东省自然科学基金项目(2014A030312013)

摘　　要：采用分批次酶解和高效液相色谱与质谱分析联用技术(HPLC-MS)对脱细胞猪小肠黏膜下层(p-SIS)基质材料中纤维连接蛋白(FN)进行了定量检测。分批次酶解消除了静态酶解时间长、酶解效率低的不足,将酶解时间从96 h缩短至6 h,降解率可以达到95%以上。采用胰蛋白酶特异性酶解得到猪源FN的一个特征多肽为SSPVVIDASTAIDAPSNLR,HPLC-MS对这一特征肽段定量检测结果表明,VIDASISTM和Biodesign两种脱细胞材料中FN的含量分别为0.43%和0.17%。方法学实验证明该方法精确度较高(RSD为1.31%)。结果表明基于质谱的分析技术可有效检测脱细胞基质中FN含量。Quantitative analysis of fibronectin in porcine small intestinal submucosa（p-SIS） acellular matrix was investigated using batch enzymatic digest and high performance liquid chromatography mass spectrometry（HPLC-MS）. The results indicated that batch enzymatic digest could improve the hydrolysis efficiency compared to static enzymatic hydrolysis. The hydrolysis time decreased to 6 hours from 96 hours,and the enzymatic hydrolysis rate amounted to 95%. Marker peptide,SSPVVIDASTAIDAPSNLR,was detected from the tryptic digest of FN. Based on the peak areas of marker peptide from FN standards with different concentration,the FN content in VIDASISTMwas determined as 0. 43%. Amount of FN in Biodesignwas determined as 0. 17%. The method precision（RSD,1. 31%） and the deviation were carried out in the methodology experiments. The results indicate the established quantitative method is effective for detecting FN content in acellular matrix.

关 键 词：猪小肠黏膜下层 脱细胞基质 纤维连接蛋白 特征肽段 液质联用技术 

分 类 号：R318.08[医药卫生—生物医学工程] R651.3[医药卫生—基础医学]