title: Expression, purification and characterization of anti-BAFF antibody secreted from the yeast Pichia pastoris.
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, pPICZalphaA, 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.
authors.affiliation: Jiangsu Province Key Laboratory for Molecular and Medical Biotechnology, Life Sciences College, Nanjing Normal University, Nanjing, 210046, China.
authors.forename: Fang
authors.initials: F
authors.lastname: Ren
authors.forename: Bao-Cun
authors.initials: BC
authors.lastname: Li
authors.forename: Na-Na
authors.initials: NN
authors.lastname: Zhang
authors.forename: Meng
authors.initials: M
authors.lastname: Cao
authors.forename: Wen-Bing
authors.initials: WB
authors.lastname: Dan
authors.forename: Shuang-Quan
authors.initials: SQ
authors.lastname: Zhang
mesh_terms: D000906:Antibodies / Q000302:isolation & purification / Q000378:metabolism; D000918:Antibody Specificity; D053264:B-Cell Activating Factor / Q000037:antagonists & inhibitors* / Q000276:immunology; D003001:Cloning, Molecular; D004797:Enzyme-Linked Immunosorbent Assay; D005434:Flow Cytometry; D005822:Genetic Vectors; D006801:Humans; D007141:Immunoglobulin Fc Fragments / Q000302:isolation & purification* / Q000378:metabolism*; D010843:Pichia / Q000276:immunology; D010957:Plasmids; D014170:Transformation, Genetic
chemical_list: D000906:Antibodies; D053264:B-Cell Activating Factor; D007141:Immunoglobulin Fc Fragments