کارآیی ترشح و گاماکربوکسیلاسیون فاکتور ix نوترکیب انسانی در سلول‌های پایدار دروزوفیلا

Abstractbackground and objectiveshuman factor ix (hfix) during maturation in the liver requires carboxylation of glutamic amino acids in the gla domain, which is involved in its secretion and activity. due to the deficiency of mammalian expression systems in the secretion and fully ?_carboxylation of recombinant coagulation factors and higher activity of ?_carboxylase enzyme in drosophila s2system, the present study was performed to evaluate ability of this system in ?_carboxylation, secretion and activity of recombinant hfix. materials and methodsin this study, following transfection of s2 cells with pmt-hfix vector, the elisa and aptt tests were used to evaluate the expression and activity of recombinant hfix. in addition, ?_carboxylation of factor ix was approved by barium citrate precipitation. the samples were analyzed on three consecutive days and being repeated three times. resultsthe coagulation results showed the secretion of active recombinant hfix by stable s2 cells. quantitative assessment of recombinant hfix in medium and cell lysis with elisa showed 94% secretion efficiency. the results of elisa on precipitated fix with barium citrate also indicated that about 45% of secreted recombinant hfix from s2 cells are fully carboxylated. conclusionsactivity and barium citrate precipitation of recombinant hfix confirmed the ability of s2 cells, unlike other insect cells, in the ?_carboxylation of factor ix. so, our results provide convincing evidence that drosophila ?_carboxylase perform necessary ?_carboxylation required for secretion and coagulation activity of recombinant hfix.