普洱茶茶褐素单克隆抗体的制备
发布时间：2018-07-27 11:13
【摘要】：普洱茶是我国云南等地区特有的一种茶类,在普洱茶加工过程中,茶褐素的含量一直增加,也是多酚氧化产物中含量最多的一类色素,并且对普洱茶的品质形成有十分重要的作用。本论文主要研究成果：以茶褐素作为抗原免疫小鼠获取脾细胞,与来源于同品系小鼠的骨髓瘤细胞SP2/0进行融合,经克隆筛选出阳性杂交瘤细胞,再将阳性杂交瘤细胞注入小鼠腹腔内制备出单克隆抗体。采用G蛋白亲和层析柱对腹水进行纯化,采用westernblot分析表明所获得的抗体为茶褐素单克隆抗体,使用ELISA法测定单抗的类型和亚型2E4杂交瘤克隆株为IgG2a型。普洱茶茶褐素单克隆抗体的获得为建立一种检测普洱茶品质方法提供一定的理论依据。
[Abstract]:Pu'er tea is a unique kind of tea in Yunnan and other regions of China. During the processing of Pu'er tea, the content of tea brownin has been increasing, and it is also the most abundant pigment in polyphenol oxidation products. And the quality of Puer tea has a very important role. The main results of this study were as follows: spleen cells were obtained from mice immunized with thefuscin as antigen and fused with myeloma cells (SP2/0) from the same strain of mice. Positive hybridoma cells were cloned and screened. Then the positive hybridoma cells were injected into the abdominal cavity of mice to prepare monoclonal antibodies. Ascites were purified by G protein affinity chromatography. Westernblot analysis showed that the obtained antibody was a monoclonal antibody to tea brownin, and the type and subtype of 2E4 hybridoma were determined by ELISA method. The preparation of monoclonal antibody against brownin in Puer tea provides a theoretical basis for the establishment of a method for determining the quality of puer tea.
【学位授予单位】：长春理工大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R392
本文编号：2147639
[Abstract]:Pu'er tea is a unique kind of tea in Yunnan and other regions of China. During the processing of Pu'er tea, the content of tea brownin has been increasing, and it is also the most abundant pigment in polyphenol oxidation products. And the quality of Puer tea has a very important role. The main results of this study were as follows: spleen cells were obtained from mice immunized with thefuscin as antigen and fused with myeloma cells (SP2/0) from the same strain of mice. Positive hybridoma cells were cloned and screened. Then the positive hybridoma cells were injected into the abdominal cavity of mice to prepare monoclonal antibodies. Ascites were purified by G protein affinity chromatography. Westernblot analysis showed that the obtained antibody was a monoclonal antibody to tea brownin, and the type and subtype of 2E4 hybridoma were determined by ELISA method. The preparation of monoclonal antibody against brownin in Puer tea provides a theoretical basis for the establishment of a method for determining the quality of puer tea.
【学位授予单位】：长春理工大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R392
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