title: Construction, purification, and characterization of anti-BAFF scFv-Fc fusion antibody expressed in CHO/dhfr- cells.
abstract: Elevated levels of B-cell-activating factor of the tumor necrosis factor family (BAFF) have been implicated in the pathogenesis of autoimmune diseases in human. In this study, we have constructed a vector for the expression of a novel compact antibody composed of anti-BAFF single-chain antibody fragment (scFv) and the Fc region (the hinge region, CH2, and CH3 domains) of human IgG1 in Chinese hamster ovary cells. The scFv-Fc fusion protein, showing spontaneous Fc fragment-mediated homodimerization via disulfide bridges, was affinity-purified on protein A Sepharose from culture supernatant. The scFv-Fc antibody was demonstrated to retain high binding affinity to antigen and prolonged clearance time in blood and to possess some human IgG crystallizable fragment effector functions such as protein A binding and antibody-dependent cellular cytotoxicity. These results suggest that this recombinant antibody may have therapeutic applications in the therapy of autoimmune disorders mediated by BAFF.
authors.affiliation: Jiangsu Province Key Laboratory for Molecular and Medical Biotechnology, Life Sciences College, Nanjing Normal University, Nanjing, Jiangsu, 210046, People's Republic of China.
authors.forename: Meng
authors.initials: M
authors.lastname: Cao
authors.forename: Peng
authors.initials: P
authors.lastname: Cao
authors.forename: Huaijiang
authors.initials: H
authors.lastname: Yan
authors.forename: Wuguang
authors.initials: W
authors.lastname: Lu
authors.forename: Fang
authors.initials: F
authors.lastname: Ren
authors.forename: Yunlong
authors.initials: Y
authors.lastname: Hu
authors.forename: Shuangquan
authors.initials: S
authors.lastname: Zhang
mesh_terms: D000818:Animals; D053264:B-Cell Activating Factor / Q000276:immunology* / Q000378:metabolism; D015153:Blotting, Western; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007141:Immunoglobulin Fc Fragments / Q000276:immunology* / Q000378:metabolism; D007128:Immunoglobulin Fragments / Q000276:immunology* / Q000378:metabolism; D008954:Models, Biological; D011993:Recombinant Fusion Proteins / Q000276:immunology* / Q000378:metabolism
chemical_list: D053264:B-Cell Activating Factor; D007141:Immunoglobulin Fc Fragments; D007128:Immunoglobulin Fragments; D011993:Recombinant Fusion Proteins