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

作者：本站编辑    发布日期：2021-03-10

Quantitative detection of fibronectin in porcine small intestinal submucosa extracellular matrix

作者：

陈毅，夏磊磊，张扬，高建萍，李赛娜，雷雄心，李勇超，张贵锋，赵博

 

中文摘要：

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，VIDASIS^TM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。

 

英文摘要：

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 VIDASIS^TM was 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

 

Keywords：

porcine small intestinal submucosa; extracellular matrix; fibronectin; marker peptide; LC-MS

 

生物学杂志，第35卷第2期，2018年4月