Preparation and characterization of monoclonal antibody specific for alpha-bungarotoxin and localization of the epitope.
We prepared a monoclonal antibody (mAb) specific for alpha-bungarotoxin (alpha-BuTX) which can neutralize the lethal toxicity of the toxin and inhibit the binding of [3H]-alpha-BuTX to the nicotinic acetylcholine receptor. The radiolabelled toxin has a high affinity for the mAb. alpha-BuTX was digested with acid protease A and the resulting peptide fragments were isolated by reverse-phase HPLC. The epitope recognized by the mAb has been localized on the basis of competition radioimmunoassay between [3H]-alpha-BuTX and the peptide fragments of alpha-BuTX towards the antibody. The epitope specific for the mAb may be located in the second loop of alpha-BuTX, probably involving residues 34-41.
Chuang
L Y
LY
Department of Biochemistry, Kaohsiung Medical College, Taiwan, R.O.C.
Lin
S R
SR
Chang
S F
SF
Chang
C C
CC
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000302:isolation & purification*; D000918:Antibody Specificity; D002038:Bungarotoxins / Q000276:immunology*; D002851:Chromatography, High Pressure Liquid; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000032:analysis*; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D009500:Neutralization Tests; D010446:Peptide Fragments / Q000032:analysis / Q000276:immunology; D011863:Radioimmunoassay; D011950:Receptors, Cholinergic / Q000302:isolation & purification
D000911:Antibodies, Monoclonal; D002038:Bungarotoxins; D000939:Epitopes; D010446:Peptide Fragments; D011950:Receptors, Cholinergic