Author/Authors :
Amiri Yekta, Amir National Institute for Genetic Engineering and Biotechnology (NIGEB),Islamic Azad University of Jahrom, ايران , Zomorodipour, Alireza National Institute for Genetic Engineering and Biotechnology (NIGEB), ايران , Khodabandeh, Mahvash National Institute for Genetic Engineering and Biotechnology (NIGEB), ايران , Daliri Chopari, Morteza National Institute for Genetic Engineering and Biotechnology (NIGEB), ايران , Kafilzadeh, Farshid Islamic Azad University of Jahrom, ايران
Abstract :
With the aim of the production of human factor VIII antigen and its corresponding antibody an epitope coding fragment of the light-chain of hFVIII, fused to a His6-tag, was isolated and over-expressed in Eschehchia coli. The over-expressed hFVIII-epitope containing peptide was confirmed by its reac- tion with a rabbit serum directed against native hFVIII as well as antiHis6-tag antibody. An expression level of 6.5 mg/l (of culture) of the C1C2-related peptide was estimated. The purified product was used to develop antibody in rabbit The immunoblotting experiment confirmed that the rabbit poly- clonal antibodies developed against the purified bacterially expressed hFVIII sub-fragment, recognizes human plasma derived FVIII. Both the produced hFVIII-related antigen and its corresponding antibody are useful in experiments using for detection and purification of hFVIII as well as the clinical diagnosis of hFVIII related disorders.
