Cloning,expression and activity of recombinant human aFGF / 中国免疫学杂志

ABSTRACT

Objective:To study the clinical application potential of aFGF and try to produce aFGF as gene engineering medicine.Methods:The complete encoding cDNA of human aFGF was isolated from human lung fibroblast with RT PCR.Recombinant human aFGF was expressed in secretory pichia expression system and purified with heparin sepharose chromatography.The activity of aFGF was detected in NIH3T3 proliferation assay,chick embryo chorioallantoic membrane assay(CAM) and wound healing assay.Results:The recombinant aFGF was expressed in large quantity(yield=12 mg/L) and was capable to promote proliferation of NIH3T3,angiogenesis and wound healing.Furthermore,those activities of aFGF could be agonized by recombinant FGFR extracellular domain.Conclusion:Recombinant human aFGF was expressed efficiently and possessed natural biological activities.