抗乙酰胆碱酯酶单克隆抗体的产生

将SP 2/O-Ag 14/小鼠骨髓瘤细胞和用乙酰胆碱脂酶免疫的Balb/c小鼠脾细胞在PEG-1000作用下进行融合,获14株分泌抗乙酰胆碱酯酶单克隆抗体的杂交瘤细胞系。用酶联免疫吸附试验(ELISA)对其分泌的抗体滴度进行了检测,其免疫鼠腹水滴度最高可达1:2 560 000。对其中5株杂交瘤分泌的滴度很高的抗体做了特异性交叉免疫试验,仅与乙酰胆碱脂酶发生特异反应,与其它4种酶(胃蛋白酶、木瓜蛋白酶、链霉蛋白酶及胰蛋白酶)无交叉反应;对其中12株杂交瘤分泌的抗体还进行了免疫球蛋白类型鉴定及染色体检查。上述杂交瘤细胞经3～6个月稳定传代后冻存于液氮中1年多后复苏,其抗体水平未见下降。

The production of anti-acetylcholinesterase monoclonal antibodies

Fourteen hybridoma cell lines excreting monoclonal antibodies directed to acetylcholinesterase (AChE)have beenestablished by fusing mouse myeloma cells and splenocytes from Balb/c mice immunized with AChE.The monoclonal antibodies produced by these hybridoma cells were identified by enzyme-linked immunosorbent assay (ELISA).Five monoclonal antibodies were examined for cross immunoactivity,all of them displayed marked specificity to AChE other than pepsin,papain,pronase and trypsin.Immunoglobulins of twelve monoclonal antibodies were classified and the chromosomes of the hybridoma cells were analyzed.Furthermore,the monoclonal antibodies excreted by three hybridoma celllines (3F3,2G8,1H11)showed significant inhibition of the activity of AChE.Hybridoma cells fairly reserved the antibody-producing capacity after stably grown for three to six months and stored in liquid nitrogen for more than one year.