Namrata Singh
Indian Institute of Chemical Biology, India
Biography
An ether extract of nine different bacterial metabolites combined with two step (ether followed by ethanol) extract of bovine bile lipid is used as an immune stimulatory drug. While characterizing the drug, we observed fibrinolytic activity in the extract through fibrinogen plate assay and fibrin zymography. Background literature emphasized major role of fibrinolytic enzymes in activating immune systems. This increased our curiosity to understand the role of these enzymes in this drug in human physiology. This fibrinolytic enzyme/s has no similarity with plasmin in terms of cross reactivity in immunoblot assay and hydrolysis of the specific substrate S-2251. In RP-HPLC analysis, the lipid extract was fractionated into several components. Interestingly, fibrinolytic activity was confined to all the fractions. To purify the enzyme, it was extracted from the lipid by aqueous buffer extraction and applied to CNBr activated fibrinogen substrate affinity column. Purified enzyme was tested for activation of complement system and wound healing through C3 binding and in-vivo wound healing assay respectively. The enzyme will be identified by mass-spectrometric analysis. Also, we propose to finger-print protein components present in bile lipid by MS analysis to have a better insight of the functionality of the lipid component of the drug.