抗人脑乙酰胆碱酯酶单克隆抗体

Monoclonal antibodies directed to human brain acetylcholinesterase

本文报道３个对人红细胞乙酰胆碱酯酶（ＡＣｈＥ）无免疫交叉反应性的抗人脑ＡＣｈＥ单克隆抗体．用纯化的人脑ＡＣｈＥ抗原多次多途径免疫ＢＡＬＢ／ｃ小鼠，取脾细胞与ＳＰ２／Ｏ－Ａｇ１４小鼠骨髓瘤细胞融合，建立了３个分泌抗人脑ＡＣｈＥ单克隆抗体（均为ＩｇＭ）的杂交瘤细胞株（Ｂ１０，Ｈ１１，Ｅ１１），小鼠腹水抗体的滴度分别为１：９６０００．１：４８０００和１：２４０００．３个抗人脑ＡＣｈＥ单克隆抗体与天然电鳐电器官ＡＣｈＥ有明显的免疫交叉反应，而与人红细胞膜ＡＣｈＥ无免疫交叉反应．３个杂交瘤细胞株经３－６个月稳定传代后冻存于液氮中，一年后复苏，细胞生长良好，分泌抗体水平未见下降．

BALB/c mice were immunizedsubcutaneously, intramuscularly and intradperitoneallyfor five times with the purified human cerebrumacetylcholinestenise(AChE).Splenocytes of the immunlzedmice were fused with SP2/O-Ag14 mouse myeloma cells.Three hybridomas(B10,N11,E11)have been developed,the dilution titers of the ascites antibodies were:96000,1:48 000 and 1:24000 respectively. The subclasses of themonoclonal antibodies and the chromosomes of hybridomacells were also determined.All the three monoclonalantibodies remarkably reacted with the native TorpedoAChE, however no obvious cross immunoreactivity couldbe detected with the humam red blood cell membraneAChE. The hybridoma cell lines were stored in liquid nitro-gen after three to six months of propagation for stability.The cells grew well and fairly reserved the antibody-se-creting capacity at resuscitation one year later.