Methods comparison of purifying anti-rat TR βΔantibodies from the antiserum
The aim of the study was to develop a procedure ofpurifying rabbit anti-rat TR β?polyclonal antibody from theantiserum. TR β?was a newly reported thyroid hormone receptorbeta isoform. To gain better understanding of this new receptor,such as its tissue distribution, its specific antibody was needed. Inthis study, chemically synthesized peptide, which is uniquelyexisted in the TR β?, was used as immunogen to generate antibodyin the rabbit. Then rabbit anti-TR β?antiserum was purified bymeans of saturated ammonium sulfate precipitation or protein Aaffinity chromatography respectively. To compare the effectivenessof these two purification methods, the purity, yield ratio andantigen binding ability of the purifications were analyzed bySDS-PAGE, protein content determination and ELISA seperately.The results showed that 98 % purity, yield of 7.47mg IgG permilliliter serum was reached via protein A affinity chromatographyand 87% purity, yield of 5.58 mg IgG per milliliter serum could beobtained by saturated ammonium sulfate precipitation. Results ofELISA analysis showed that IgG purified by either method couldbind to recombinant TR βΔprotein with same affinity. So, we drewthe conclusion that the protein A affinity chromatography methodwas more suitable for the purification of the anti-TR βΔpolyclonalantibody from the antiserum.
Ying Liu Bei Sun Yan Ma Liling Hu Xueqin Zhao Dongchun Liang Aijun Zuo Gang Guo JingyuZhang
Institute of Endocrinology and Metabolic Disease Hospital, Tianjin Medical University Key lab of Hor Institute of Endocrinology and Metabolic Disease Hospital, Tianjin Medical University Key lab of Hor
国际会议
成都
英文
1-3
2010-06-18（万方平台首次上网日期，不代表论文的发表时间）