Bacterial Expression and Purification of C1C2 Domain of Human Factor VIII
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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 Escherichia coli. The over-expressed hFVIII-epitope containing peptide was confirmed by its reaction 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. Theimmunoblotting experiment confirmed that the rabbit polyclonal antibodies developed against the purified bacteriallyexpressed hFVIII sub-fragment, recognizes human plasmaderived FVIII. Both the produced hFVIII-related antigen andits corresponding antibody are useful in experiments usingfor detection and purification of hFVIII as well as the clinicaldiagnosis of hFVIII related disorders.
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Journal title
volume 4 issue 2
pages 104- 111
publication date 2006-04-01
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