Purification and characterization of platelet aggregation inhibitor from the venom of Bitis arietans

Disintegrins are the antagonists of integrin receptors that can be found mostly in snakes’ venom. They can inhibit platelet aggregation, thus preventing the formation of blood clots. By blocking the integrin receptors of cancer cells, disintegrins can inhibit proliferation and metastasis. Thus, the search for new sources of disintegrins and development of methods of their purification is an important task of modern biotechnolo­gy. This work was dedicated to the purification and characterization of inhibiting polypeptides from Bitis arietans­ venom. Crude venom of B. arietans was fractionated using ion-exchange chromatography on Q Sepharose followed by size-exclusion chromatography on Superdex 75 using FPLC method. Analysis of molecular weight of protein components was performed using SDS-PAGE and MALDI-TOF analysis on Voyager-DE. Aggregation of platelet-rich plasma (PRP) in the presence of platelet aggregation inhibitor was investigated using aggregometry on the AR2110. MTT test was used for measuring HeLa cells proliferation and survival in vitro. Two-step chromatography allowed us to obtain fraction that contained polypeptides possessing the dose-dependent inhibitory action on ad