Amidolytic Activity of Factor VII Expressed in Iranian Lizard Leishmania

Abstract

Over many years, variable gene expression systems have been used obtaining of Factor VII protein, but each one has certain limitations. Therefore, the main goal of this study was to assess the biological activity of purified and recombinant factor VII expressed in Iranian lizard Leishmania. After transferring recombinant construct containing FVII gene to Leishmania, first, the expression of 55 kDa FVII protein in transfected cells was confirmed by analyzing cell lysate using SDS-PAGE and Western Blotting techniques. Then, FVII was purified by NI-NTA-His Tag- resin through an affinity chromatography. The chromogenic activity of human recombinant Factor VII (amidolytic activity) in supernatant and pellet fractions of Leishmania promastigotes was done using ELISA method. The amidolytic activity of rhFVII was about 0.0125 IU/ml in the concentrated cell sediment and 0 IU/ml in the supernatant in the first 60 minutes and after that time, none of the samples showed acceptable activity. HIGHLIGHTS • Factor VII has important role in the treatment of coagulation and hemophilic disease. • Recombinant FVII was expressed in Iranian Lizard Leishmania. • The amidolytic activity of rhFVII was about 0.0125 IU/ml in cell pellet. • No amildolytic activity was observed in the supernatant.

Keywords

• amidolytic activity
• chromogenic assay
• lizard leishmania
• recombinant factor vii