酚酞单克隆抗体的制备
酚酞单克隆抗体的制备
目的建立免疫学快速检测减肥类保健食品中非法添加的酚酞, 制备酚酞单克隆抗体并进行评价。方法利用碳二亚胺(carbodiimide, EDC)法合成免疫原和包被原, 用免疫原免疫Balb/C小鼠, 取小鼠脾脏与SP2/0鼠骨髓瘤细胞融合。采用竞争结合双阳性两步筛选法, 筛选出能分泌特异性抗体的杂交瘤细胞, 利用有限稀释亚克隆方法得到单株细胞; 采用体内诱生法制备腹水型单克隆抗体。利用辛酸-饱和硫酸铵法对腹水型抗体进行纯化, 利用酶联免疫吸附法鉴定纯化后的抗体。结果成功合成了酚酞-BSA免疫原和酚酞-OVA包被原, 筛选获得酚酞杂交瘤细胞株FT/BSA/2019, 单克隆抗体的效价1×105。结论本研究初步建立了特异性高的酚酞单克隆抗体的制备方法。
ObjectiveTo establish immunological rapid detection of illegally added phenolphthalein in dietetic health foods, and to prepare and evaluate phenolphthalein monoclonal antibodies. MethodsThe immunogen and coating antigen were synthesized using a carbodiimide (EDC) method. Balb/C mice were immunized with the synthetic immunogen, and the mouse spleen was fused with SP2/0 mouse myeloma cells. The hybridoma cells capable of secreting specific antibodies were screened by adopting a competition and double-positive two-step screening method, individual cell line was obtained by using a limited dilution subcloning method, and the ascites-type monoclonal antibody was prepared by adopting an in-vivo induction assay. The ascites-type antibody was purified by caprylic acid-saturated sulfuric acid method, and the purified antibody was identified by enzyme-linked immunosorbent assay (ELISA). ResultsPhenolphthalein -BSA immunogen and phenolphthalein-OVA encapsulant were successfully synthesized, FT/BSA/2019 of phenolphthalein hybrid tumor cell line was screened, and the titer of monoclonal antibody was 1×105. Conclusion This paper establish the preparation method of the specific high-specificity phenolphthalein monoclonal antibody.
标题：酚酞单克隆抗体的制备
英文标题：Preparation of monoclonal antibody of phenolphthalein illegally added in health products
作者：
谭攀 深圳市三方圆生物科技股份有限公司
吴鑫 深圳市三方圆生物科技股份有限公司
蔡杜曼 广东环境保护工程职业学院
胡金梅 广东环境保护工程职业学院
中文关键词:酚酞,单克隆抗体,酶联免疫吸附法,
英文关键词:phenolphthalein,monoclonal antibody,enzyme-linked immunosorbent assay,
发表日期：2019-11-03
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