抗α-银环蛇毒素单克隆抗体的制备、鉴定及表位定位

Preparation and characterization of monoclonal antibody specific for alpha-bungarotoxin and localization of the epitope.

作者信息

Chuang L Y, Lin S R, Chang S F, Chang C C

机构信息

Department of Biochemistry, Kaohsiung Medical College, Taiwan, R.O.C.

出版信息

Toxicon. 1989;27(2):211-9. doi: 10.1016/0041-0101(89)90134-7.

DOI:10.1016/0041-0101(89)90134-7

PMID:2470166

Abstract

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.

摘要

我们制备了一种针对α-银环蛇毒素（α-BuTX）的单克隆抗体（mAb），它可以中和该毒素的致死毒性，并抑制[3H]-α-BuTX与烟碱型乙酰胆碱受体的结合。放射性标记的毒素对该单克隆抗体具有高亲和力。用酸性蛋白酶A消化α-银环蛇毒素，所得肽片段通过反相高效液相色谱法分离。基于[3H]-α-BuTX与α-银环蛇毒素肽片段对该抗体的竞争放射免疫测定，确定了该单克隆抗体识别的表位。该单克隆抗体特异的表位可能位于α-银环蛇毒素的第二个环中，可能涉及34-41位氨基酸残基。