识别脊神经感觉神经元单克隆抗体的制备

THE PREPARATION OF MONOCLONAL ANTIBODY RECOGNIZING THE SENSORY NEURON OF SPINAL NERVE

将脊神经感觉神经元特有的蛋白组份作为抗原,注入BABL/c小鼠腹腔中。采集经免疫的小鼠脾细胞,以聚乙二醇(PEG)为促融剂,使之与小鼠骨髓瘤细胞SP2/0融合。应用HAT选择性培养液培养杂交瘤细胞。三周后,以ABC法检测,筛选出仅对感觉神经元起反应的Ⅱ6H杂交瘤细胞株,三次有限稀释,克隆化后,体外培养达一年之久,仍能稳定地分泌抗体。该抗体属IgG_1亚类,特异性地识别感觉神经元特有的蛋白组份。应用该单抗,可鉴别周围神经干中的感觉与运动纤维,亦可研究感觉神经的发育以及结构与功能的关系。

As an antigen, the crude abstract of the sensory neuronal protein was iniected intraperitoneally to BABL/c mouse. The immunized spleen ceils were fused with SP2/0 cells using PEG as fusion promoting agent. The hybridoma cells were suspended in HAT selective cultures and the positive reaetion to sensory neurons were detected by ABC method 3 weeks later, the individual hydridized cell showing positive reaction only to sensory neurons were selected and isolated. After cloning three times, the cells cultivated in vitro for one year could still secrete antibody steadily. It belonged to IgG_1 and was specific for the sensory neuron protein. The monoclonal antihody might be used to identify sensory from motor fibres, and to study the development of the sensory neurons and the relations between structure and function.