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Expression,purification and characterization of anti-BAFF antibody secreted from the yeast Pichia pastoris
Authors:Fang Ren  Bao-Cun Li  Na-Na Zhang  Meng Cao  Wen-Bing Dan  Shuang-Quan Zhang
Affiliation:(1) Jiangsu Province Key Laboratory for Molecular and Medical Biotechnology, Life Sciences College, Nanjing Normal University, Nanjing, 210046, China
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.
Keywords:B-cell activating factor  Binding activity  Heterologous expression   Pichia pastoris   scFv-Fc
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