Expression,purification and characterization of anti-BAFF antibody secreted from the yeast Pichia pastoris |
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Authors: | Fang Ren Bao-Cun Li Na-Na Zhang Meng Cao Wen-Bing Dan Shuang-Quan Zhang |
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Affiliation: | (1) Jiangsu Province Key Laboratory for Molecular and Medical Biotechnology, Life Sciences College, Nanjing Normal University, Nanjing, 210046, China |
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Abstract: | ![]() B-Cell activating factor (BAFF) is critical for B cell survival and maturation; excessive expression of it corrupts B-cell tolerance and may lead to autoimmunity. The gene, scFv-Fc, coding for the antibody of BAFF was inserted into the eukaryotic expression vector, pPICZαA, and transformed into Pichia pastoris. A high-level expression strain was obtained using a ‘yeastern blotting’ method. The scFv-Fc antibody was purified and 56 mg was obtained from 1 l of culture supernatant. It retained high binding activity to both soluble BAFF and membrane-bound BAFF. |
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Keywords: | B-cell activating factor Binding activity Heterologous expression Pichia pastoris scFv-Fc |
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