Characterization of an anti-thioredoxin monoclonal antibody.
An anti-E. coli thioredoxin monoclonal antibody, IMM-3C6, which showed high specificity to thioredoxin as assessed by indirect ELISA, was generated using hybridoma technology. The affinity constant of IMM-3C6 to thioredoxin was 0.40 x 10(9) M: (-1) and its sensitivity to thioredoxin fusion protein in dot blotting was 50 ng. In sandwich ELISA, it detected thioredoxin fusion protein between 16 and 150 ng/ml. By using IMM-3C6 as the ligand, thioredoxin fusion protein was successfully purified by affinity chromatography. IMM-3C6 was confirmed to be a useful tool for immunoassay and purification of thioredoxin fusion proteins.
Zhang
Wei
W
Institute of Molecular Medicine, Nanjing University, Nanjing, 210093, China.
Zhang
Wang
W
Tang
Yanchun
Y
Zhang
Jing
J
Liu
Jian-Ning
JN
D000818:Animals; D000911:Antibodies, Monoclonal / Q000302:isolation & purification*; D004797:Enzyme-Linked Immunosorbent Assay; D029968:Escherichia coli Proteins / Q000276:immunology* / Q000302:isolation & purification; D005260:Female; D051379:Mice; D008807:Mice, Inbred BALB C; D011993:Recombinant Fusion Proteins / Q000302:isolation & purification; D013879:Thioredoxins / Q000276:immunology*
D000911:Antibodies, Monoclonal; D029968:Escherichia coli Proteins; D011993:Recombinant Fusion Proteins; D013879:Thioredoxins