Expression, purification and characterization of anti-BAFF antibody secreted from the yeast Pichia pastoris.
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.
Ren
Fang
F
Jiangsu Province Key Laboratory for Molecular and Medical Biotechnology, Life Sciences College, Nanjing Normal University, Nanjing, 210046, China.
Li
Bao-Cun
BC
Zhang
Na-Na
NN
Cao
Meng
M
Dan
Wen-Bing
WB
Zhang
Shuang-Quan
SQ
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
D000906:Antibodies; D053264:B-Cell Activating Factor; D007141:Immunoglobulin Fc Fragments