乙酰胆碱受体单链噬菌体抗体的筛选与鉴定

Selection and analysis of single chain antibodies against acetylcholine receptor from phage antibody library

筛选乙酰胆碱受体（ＡＣｈＲ）单链抗体（ＳｃＦｖ），制备可溶性ＡＣｈＲＳｃＦｖ以预防、治疗和诊断重症肌无力。采用电鳐电器官的ＡＣｈＲ（ｔＡＣｈＲ）以亲和吸附法，从鼠源性噬菌体ＳｃＦｖ抗体库中筛选出ＡＣｈＲ特异性抗体。以ＥＬＩＳＡ鉴定噬菌体抗体的特异性，以免疫荧光技术鉴定噬菌体抗体对人肌肉冰冻切片中ＡＣｈＲ的结合特异性。经４轮ＡＣｈＲ筛选后，获得了能与ＡＣｈＲ结合的噬菌体抗体，这种抗体也能与人肌肉冰冻切片中的ＡＣｈＲ结合。以ｔＡＣｈＲ为抗原对噬菌体抗体库进行亲和吸附筛选，可以得到ＡＣｈＲＳｃＦｖ，并且这种抗体也能与人肌肉冰冻切片中ＡＣｈＲ特异性结合。

Objective:To make single chain antibodies(ScFv) against acetylcholine receptor(AChR) which could probably be used in prevention,treatment and diagnosis of myasthenia gravis.Methods:To select AChR ScFv from phage antibody library constructed from unimmunized mouse spleen cells against antigen Torpedo AChR(tAChR).The binding activities of AChR ScFv to tAChR and AChR in cryosections of human muscle(hAChR) were determined by ELISA and immunohistochemical staining technique.Results:AChR ScFv were obtained from the phage antibody library after four round panning.One AChR ScFv was found to be able to bind hAChR in cryosection of muscle.Conclusions:It is possible to make AChR ScFv from the phage antibody library by panning against AChR.This will allow us to make soluble AChR ScFv to block pathogenic AChR antibodies in myasthenia gravis.