Recombinant Expression,Purification and Activity of Complement C5 Inhibitor OmCI
Ornithodoros moubata complement inhibitor(OmCI,1)is a complement C5 inhibitor for the treatment of complement-mediated diseases.In this study,a recombination,expression,isolation and purification method of 1 was established,and activity of the obtained 1 was verified.The protein was expressed in extracellular soluble form by high density fermentation technique of recombinant Escherichia coli(E.coli),and the yield of fusion protein was 317 mg/L.The fermentation supernatant was pretreated by acid-adjusted clarification,enzyme digestion to remove the tagged proteins.Then it was purified by anion-exchange chromatography and reversed-phase chromatography to get the target protein with a purity of 98.2％,and the total yield was 62.8％.The relative molecular weight of target protein was verified to be correct by mass spectrometry.According to the results of the in vitro complement classical pathway inhibition assay,the IC50 and the binding constant(KD)to complement C5 were 8.59 and 4.74 nmol/L,respectively.The results proved that 1 was successfully expressed in E.coli and exhibited C5 inhibitory activity.This study provides a reference for the efficient preparation of complement C5 inhibitors.
Ornithodoros moubata complement inhibitor(OmCI)recombinant expressionfermentationpurificationactivity test
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