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| High Level Expression of Human ScFv against bFGF in Pichia pastoris |
| LV Wei-dong1, DU Chao-chao1, WANG Hong1, JIANG Hao-wu1, LAO Xue-jun2, SONG Qi-fang1, DENG Ning1 |
1. Guangdong Province Key Laboratory of Molecular Immunology and Antibody Engineering, Jinan University, Guangzhou 510632, China;
2. Department of General Surgery, The First Affiliated Hospital of Jinan University, Guangzhou 510632, China |
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Abstract In order to high level express anti-bFGF ScFv antibody in Pichia pastoris, the gene of human anti-bFGF ScFv was subcloned, and constructed into expression vector pPICZαA. The constructed expression vector pPICZαA-ScFv was linearised and transformed to Pichia pastoris by electroporation. The transformants were induced by methanol, and the anti-bFGF ScFv was expressed. The expression products were purified by affinity chromatography of Ni-Seproase 6 FF and ion exchange chromatography of DEAE Sepharose FF. The results of SDS-PAGE and Western blotting showed that anti-bFGF ScFv was high level expressed successfully and the expression quantity was about 124mg/L. The target protein was purified from the expression products and the purity was more than 95 %. The results of ELISA showed that the purified recombinant ScFv could combine with bFGF specifically. The results of CCK8 showed that the purified recombinant ScFv could inhibit the proliferation of A549 cells in a dose-dependent manner in vitro. The results demonstrated that the anti-bFGF ScFv can be high level expressed in Pichia pastoris with good biological activity.
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Received: 05 January 2012
Published: 25 May 2012
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