Expression and Purification of Recombinant Human Serum Albumin in Hansenula polymorpha

China Biotechnology

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Expression and Purification of Recombinant Human Serum Albumin in Hansenula polymorpha

fan-hong meng

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Abstract Based on Hansenula polymorpha's genetic code frequency meter, the gene of optimized codon human serum albumin (HSA) was ligated with vector of Hansenula polymorpha by T4 DNA ligase, after EcoR I and Hind III double digestion, to construct recombinant HSA (rHSA) expression vector. Transformed to Hansenula polymorpha strain by electroporation, the transformants were incubated on MD medium to select URA3+ phenotype. The highly expressive strain was selected by the methods of SDS-PAGE and Western-blotting. Fermented in feed-batch 30L-fermenter, about 1.033g/L of rHSA was obtained, according to the result of ELISA. The proteolytic degradation of rHSA turned obviously after 61h of induction and the concentration of rHSA decreased to less in 150h after induction. The rHSA was purified by streamline SP exchange chromatography、phenyl Bio-sep 6FF hydrophohic interaction chromatography and DEAE sepharose exchange chromatography, and electrophoresis-grade HSA could be obtained. Diluting the purified HAS, we immunized several group of mice and got anti-serum. Checking mouse's immunogenity by double agar diffusion test, HSA and rHSA were demonstrated with the similarly reactive character of antibody and antigen.

Key words： Hansenula polymorpha Purification Human syrum albumin

Received: 19 September 2006 Published: 25 December 2006

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fan-hong meng . Expression and Purification of Recombinant Human Serum Albumin in Hansenula polymorpha. China Biotechnology, 2006, 26(12): 11-17.

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https://manu60.magtech.com.cn/biotech/ OR https://manu60.magtech.com.cn/biotech/Y2006/V26/I12/11

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