Fibronectin Purification and Fragmentation
Fibronectin, an extra cellular matrix glycoprotein was purified from human plasma obtained from Regional Cancer Centre Trivandrum. Gelatin based affinity chromatography was used for the purification of the protein. The usage of pressure was found to have prominent impact in the purification of the protein by reducing albumin contamination, which we suspect would have non specifically bound to the gelatin resin. The quality of the protein purified was analysed using SDS –PAGE. The protein sample was further dialysed to remove urea as our future experiment required pure Fibronectin.
An experiment was also done to develop angiogenesis assay model using the chorioallantoic membrane (CAM) of fertilized chicken egg. Dimethyl sulfoxide (DMSO) was found to inhibit angiogenesis and sodium nitroprusside was found to have vasodilation effect, validating our assay model. We also tried to purify fibronectin from zebrafish using two different concentrations of urea which would solubilise the insoluble cellular fibronectin.