Abstract
Bovine pericardium (BP) is an important biomaterial used in the production of glutaraldehyde‐fixed heart valves and tissue‐engineering applications. The ability to perform proteomic analysis on BP is useful for a range of studies, including investigation of immune rejection after implantation. However, proteomic analysis of fibrous tissues such as BP is challenging due to their relative low‐cellularity and abundance of extracellular matrix. A variety of methods for tissue treatment, protein extraction, and ;fractionation were investigated with the aim of producing high‐quality 2‐DE gels for both water‐ and lipid‐soluble BP proteins. Extraction of water‐soluble proteins with 3‐(benzyldimethylammonio)‐propanesulfonate followed by n‐dodecyl β‐D‐maltoside extraction and ethanol precipitation for lipid‐soluble proteins provided the best combination of yield, spot number, and resolution on 2‐DE gels (Protocol E2). ESI‐quadrupole/ion trap or MALDI‐TOF/TOF MS protein identifications were performed to confirm bovine origin and appropriate subcellular prefractionation of resolved proteins. Twenty‐five unique, predominantly cytoplasmic bovine proteins were identified from the water‐soluble fraction. Thirty‐two unique, predominantly membrane bovine proteins were identified from the lipid‐soluble fraction. These results demonstrated that the final protocol produced high‐quality proteomic data from this important tissue for both cytoplasmic and membrane proteins.