抗鱼卵透明带单克隆抗体淋巴细胞杂交瘤的建立及初步鉴定

Establishment and Primary Identification of Hybridoma of Anti- zona Monoclonal Antibody

详细介绍了抗鱼（花鲢Anstichthysnobilis）卵透明带单克隆抗体淋巴细胞杂交瘤的制备及鉴定方法，用佛氏佐剂乳化热溶花鲢鱼卵透明带，然后免疫雌性BALB／C小鼠，取其脾细胞与骨髓瘤Sp2／0融合产生杂交瘤，经3次再克隆后，用间接ELISA方法筛选出一株抗鱼卵透明带的单克隆抗体杂交瘤，经免疫双扩散试验测得该种单克隆抗体为IgG1，经过染色体检查知其染色体数目在86～108之间，为93.6±3.81，杂交瘤细胞染色体经检查发现有中间和亚中着丝点染色体，符合杂交细胞染色体特征。

The present investigation has been carried out to produce and characterize the anti - zona monoclonal antibody against zona pellucide antigen of Aristichtys nobilis. Female BALB/C mouse were immunized by heat solubilized zona pellucide antigen emulsified in equal amount of freund adjuvant. The spleen cells were fused with SP2/0 myeloma cells through the application of routine techniques with some modifications. Modified procedures have been proved to be successful, percentage of fusing rate can ove r90% and about 10% positive rate can be Obtained. One hybridoma cell line has been identified to produce specific monoclonal antibody by using indirect ELISA. Double immunal diffusion analysis has been shown that the antibody belongs to IgGI subgroup. Specific visalization method indicates that the chromosome numbers are scaled between 86 - 108, averaged 93. 6 ± 3. 81 (x ± SD).