title: Expression and purification of recombinant human serum albumin fusion protein with VEGF165b in Pichia pastoris.
abstract: VEGF165b is an endogenous transcriptional splice variant of VEGF and has been shown to have a therapeutic potency as an anti-cancer agent. In this report, a fusion gene consisting of a human VEGF165b and a human albumin (HSA) gene was constructed and then inserted into a pPIC9k vector. The recombinant fusion protein, rhHSA-VEGF165b, was over expressed in the methylotrophic yeast Pichia pastoris under the control of AOX1 promoter. After induction with methanol, the expression level of rhHSA-VEGF165b was 275 mg/L in broth. The fusion protein rhHSA-VEGF165b was purified to more than 95% purity by using Blue Sepharose Fast Flow and SP Sepharose Fast Flow. Biological activity of the prepared rhHSA-VEGF165b was characterized by transwell migration assay, retaining about 9% of that of unmodified rhVEGF165b on a molar basis. Data from mice show that the serum half-life time of rhHSA-VEGF165b was nearly 20 times longer than that of rhVEGF165b.
authors: Molecular Pharmacol Laboratory, Sch Med and Pharmaceut, Jiangnan University, Wuxi 214122, PR China. ry_zhu@sina.com
authors: Rui-Yu
authors: RY
authors: Zhu
authors: Xin
authors: X
authors: Xin
authors: Hui-yun
authors: HY
authors: Dai
authors: Qi
authors: Q
authors: Li
authors: Jian-Yong
authors: JY
authors: Lei
authors: Yun
authors: Y
authors: Chen
authors: Jian
authors: J
authors: Jin
mesh_terms: D000818:Animals; D002465:Cell Movement / Q000187:drug effects; D003001:Cloning, Molecular / Q000379:methods; D005822:Genetic Vectors / Q000235:genetics; D061307:Human Umbilical Vein Endothelial Cells / Q000187:drug effects; D006801:Humans; D051379:Mice; D010843:Pichia / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000097:blood / Q000235:genetics* / Q000302:isolation & purification* / Q000494:pharmacology; D012709:Serum Albumin / Q000032:analysis / Q000235:genetics* / Q000302:isolation & purification* / Q000494:pharmacology; D015854:Up-Regulation; D042461:Vascular Endothelial Growth Factor A / Q000097:blood / Q000235:genetics* / Q000302:isolation & purification* / Q000494:pharmacology
chemical_list: D011993:Recombinant Fusion Proteins; D012709:Serum Albumin; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A