Expression and purification of recombinant human serum albumin fusion protein with VEGF165b in Pichia pastoris

  • Zhu R
  • Xin X
  • Dai H
 et al. 
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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. © 2012 Elsevier Inc. All rights reserved.

Author-supplied keywords

  • Expression
  • Human serum albumin
  • Pichia pastoris
  • Purification
  • VEGF165b

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Authors

  • R.-Y. Zhu

  • X. Xin

  • H.-Y. Dai

  • Q. Li

  • J.-Y. Lei

  • Y. Chen

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