新型游离三碘甲腺原氨酸人工抗原的合成与表征

doi:10.3724/SP.J.1095.2013.20570

摘要/Abstract

摘要： 利用快速、温和的“Click Chemistry(点击化学)”反应，在游离三碘甲腺原氨酸(FT3)的羧基上引入一个带有三唑环的羧基“间接臂”，生成半抗原(FT3C)。再用N-羟基琥珀酰亚胺活性酯法，将半抗原(FT3C)分别与牛血清白蛋白(BSA)和钥孔戚血蓝素(KLH)偶联，合成了免疫抗原(FT3C-BSA) 和包被抗原(FT3C-KLH)。免疫抗原(FT3C-BSA)的紫外吸收光谱相对于FT3C和BSA有明显差异，表明成功实现了偶联，经计算得到偶联比为7∶1；用FT3C-BSA免疫Balb/c小鼠，获得高效价的抗FT3血清。采用间接ELISA法检测抗体的IC50值为0.37 mg/L,抗血清效价可达6.24×104，是制备FT3人工抗原的一种新方法。

关键词: 游离三碘甲腺原氨酸, 点击化学, 半抗原, 人工抗原

Abstract: FT3C hapten was synthesized via a fast, mild “Click Chemistry”, which contained a “spacer arm” of triazole ring conjugated to carboxyl site of free triiodothyronine(FT3). Immunogen and coating antigen of FT3C were respectively prepared by the coupling reaction between hapten and bovine serum albumin(BSA) or keyhole limpet hemocyanin(KLH), using the N-hydroxysuccinimide active ester method. UV spectrum of the artificial antigen(FT3C-BSA) differed significantly from those of hapten(FT3C) and BSA, the results also showed that FT3C was conjugated to BSA successfully, the coupling ratio of hapten to BSA was 7∶1; the antibody with high titer（6.24×104） and IC50 value of 0.37 mg/L was produced through immunization of Balb/c mice. This study provided a new way to prepare FT3 artificial antigen.

Key words: free triiodothyronine, Click Chemistry, hapten, artificial antigen

中图分类号:

O629.8

印晓梅, 于海涛, 王小龙. 新型游离三碘甲腺原氨酸人工抗原的合成与表征[J]. 应用化学, DOI: 10.3724/SP.J.1095.2013.20570.

YIN Xiaomei, YU Haitao*, WANG Xiaolong . Synthesis and Characterization of a New Artificial Free Triiodothyronine Antigen[J]. Chinese Journal of Applied Chemistry, DOI: 10.3724/SP.J.1095.2013.20570.

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