Prokaryotic Expression, Purification and Polyclonal Antibody Preparation of Human Heart Type Fatty Acid Binding Protein

Objective We aimed to prepare purification H-FABP recombinant protein for antibody preparation and functional analysis.Method The encoding sequence of human H-FABP was amplified with RT-PCR and cloned into plasmid pET28a to establish the prokaryotic expressing system.E.coli BL21 was transformed by the recombinant plasmid pET28a-H-FABP.Expression of the recombinant protein was induced with IPTG and purified by Q Sepharose F.F.anion chromatography column.The recombinant H-FABP was assayed by using Immuno-colloidal gold method and Western blotting.Result SDS-PAGE analysis suggested the recombinant protein was expressed as inclusion body,with molecular weight of about 15kD,which consistent with the expecting value.After washing,the recombinant protein was purified by affinity chromatography and the purity coefficient was up to 95%.Conclusion Purified human H-FABP recombinant protein prepared in this study will facilitate antibody preparation,structure investigation and protein functional analysis.