Enrichment of the antibodies against the C-terminus of Taiwan cobra cobrotoxin using dimeric glutaraldehyde-modified toxin as an immunogen.
The repertoire of antibodies producing by immunizing rabbits with cobrotoxin and dimeric glutaraldehyde-modified cobrotoxin (dGA-cobrotoxin) was analyzed by studying the immunoreactivity of the two antibody preparations toward cobrotoxin, GA-cobrotoxin and recombinant cobrotoxin. The results of enzyme-linked immunoassay revealed that the two antibody preparations exhibited a higher reactivity against their cognate antigen. Moreover, different behavior was observed for the reactivity of the two antibody preparations against GA-cobrotoxin and recombinant cobrotoxin. Notably, distortion of disulfide linkages at the C-terminus resulted in a reduced decrease in the antigenic activity of recombinant cobrotoxin toward anti-cobrotoxin antibodies compared to anti-dGA-cobrotoxin antibodies. Affinity purification of the antibodies against the C-terminus of cobrotoxin revealed that its amount represented 77% and 35.5% of the total anti-dGA-cobrotoxin antibodies and the total anti-cobrotoxin antibodies, respectively. These findings suggest that the antibody preparation elicited by dGA-cobrotoxin enriches the content of antibodies recognizes the C-terminal region of native cobrotoxin.
Chang
Long-sen
LS
Institute of Biomedical Sciences, National Sun Yat-Sen University, 804, Kaohsiung, Taiwan, ROC. lschang@mail.nsysu.edu.tw
Lin
Rose
R
Chen
Ku-chung
KC
Chang
Chun-chang
CC
D000818:Animals; D000915:Antibody Affinity; D000916:Antibody Diversity; D000997:Antivenins / Q000145:classification / Q000276:immunology*; D003039:Cobra Neurotoxin Proteins / Q000737:chemistry / Q000276:immunology*; D004220:Disulfides; D017815:Elapidae / Q000502:physiology; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes; D005976:Glutaral / Q000737:chemistry / Q000276:immunology*; D011817:Rabbits; D011994:Recombinant Proteins / Q000737:chemistry / Q000276:immunology; D013624:Taiwan; D014614:Vaccines, Synthetic
D000997:Antivenins; D003039:Cobra Neurotoxin Proteins; D004220:Disulfides; D000939:Epitopes; D011994:Recombinant Proteins; D014614:Vaccines, Synthetic; D005976:Glutaral