Quantitative Detection of Fibronectin in Porcine Small Intestinal Submucosa Extracellular Matrix
by Publications from Biosis Healing
By:
CHEN Yi, XIA Lei-lei, ZHANG Yang, GAO Jian-ping, LI Sai-na, LEI Xiong-xin, LI Yong-chao, ZHANG Gui-feng, ZHAO Bo
(1. Beijing Biosis Healing Biological Technology Co., Ltd., Beijing 100026; 2. State Key Laboratory of
Biochemical Engineering, Institute of Process Engineering, CAS, Beijing 100190; 3. School of Life
Science and Technology, Henan Institute of Science and Technology, Xinxiang 453003, China)
Abstract Quantitative analysis of fibronectin in porcine small intestinal submucosa (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 R 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.
Keywords porcine small intestinal submucosa; extracellular matrix; fibronectin; marker peptide; LC-MS
