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| Construction and Expression of sGLP-1- HSA Fusion protein |
| GAO Hua1 XU Chong-bo1 LI Yuan2 |
| 1.Key Laboratory of Bioorganic Chemistry of Liaoning Province|College of Medicine| Dalian University|Dalian116622|China 2.Dalian D.N Bioengineering CO．|LTD|Dalian116600|China |
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Abstract To reduce the clearance of sGLP-1, sGLP-1-HSA fusion protein was constructed by the fusion of the C-terminus of sGLP-1 to the N-terminus of HSA via an 6 amino acids linker and expressed in the pichia pastoris.Screen the strain with resistsnce of 4g/L G418. After fermentation in a 5L bioreactor，the expression of sGLP-1- HSA was induced by methanol.After 48h, the expression level reached 0.8g/L.The sGLP-1- HSA was abtained following a series of purification steps such as ultrafiltration, sephadex G-25 and blue- sepharose. The purity of the fermentation product could reach 95% by means of HPLC，and the total recovery yield could reach 60%.The biological activity of sGLP-1- HSA in GKⅡ diabetes rat and healthy rat suggested that the plasma glucose level was significantly lower after subcutaneous administration of sGLP-1- HSA..Expression of sGLP-1- HSA fusion protein can provide foudation for abtaining a larger quantity of recombinant sGLP-1- HSA for experimented and clinic studies.
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Received: 19 February 2009
Published: 28 July 2009
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