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(FT3CKLH)。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。
FT3 C 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 FT3 C 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（ FT3 C-BSA） differed significantly from those of hapten （FT3 C ） and BSA, the results also showed that FT3 C was conjugated to BSA successfully, the coupling ratio of hapten to BSA was 7 ： 1 ; the antibody with high titer （ 6. 24 × 10^4 ） and ICs0 value of 0. 37 mg/L was produced through immunization of Balb/c mice. This study provided a new way to prepare T3 artificial antigen.
出处
《应用化学》
CAS
CSCD
北大核心
2013年第11期1281-1284,共4页
Chinese Journal of Applied Chemistry
基金
江苏省自然科学基金(BK2011813)
