| 25-羟基维生素D3人工完全抗原的合成与鉴定 |
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| 作者姓名： | Wan WW Zhang HJ Ye HM Wan LQ Cai LL Zheng LM Zeng CM |
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| 作者单位： | 1. 厦门大学生物医学研究院转化医学中心,福建厦门,361005
2. 南昌大学第一附属医院普三科,江西南昌,330006 |
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| 摘 要： | 目的:合成25-羟基维生素D3人工完全抗原,并制备抗25-羟基维生素D3的特异性抗体。方法:将25-羟基维生素D3进行化学修饰加入羧基活性基团,合成具有半抗原结构特征的25-羟基维生素D3-半琥珀酸酯。采用碳二亚胺法,将25-羟基维生素D3-半琥珀酸酯,分别与牛血清白蛋白(BSA)和卵清蛋白(OVA)偶联,合成人工完全抗原25-羟基维生素D3-半琥珀酸酯-BSA和25-羟基维生素D3-半琥珀酸酯-OVA。通过紫外吸收光谱,SDS-PAGE和MALDI-TOF进行偶联鉴定。用25-羟基维生素D3-半琥珀酸酯-BSA免疫小鼠,获得抗25-羟基维生素D3抗体免疫血清。结果:25-羟基维生素D3-半琥珀酸酯与BSA的偶联比为(12±0.16)∶1,免疫小鼠后获得高效价(效价为6.25×10-4)的抗体,且标准品浓度在37.5～600 ng/mL范围具有显著的竞争性线性关系,检测的灵敏度为37.5 ng/mL。结论:成功合成了25-羟基维生素D3人工完全抗原,制备出25-羟基维生素D3的抗体且其线性关系显著,灵敏度较高,为进一步研制检测25-羟基维生素D3的试剂盒奠定了基础。
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| 关 键 词： | 25-羟基维生素D3 半抗原-[C-3]-载体蛋白偶联 人工完全抗原 |
Synthesis and identification of artificial complete antigen 25-hydroxyvitamin D(3); |
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| Wan WW,Zhang HJ,Ye HM,Wan LQ,Cai LL,Zheng LM,Zeng CM.Synthesis and identification of artificial complete antigen 25-hydroxyvitamin D(3);[J].Journal of Cellular and Molecular Immunology,2011,27(11):1173-1175. |
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| Authors: | Wan Wei-wei Zhang Huan-jing Ye Hui-ming Wan Li-qin Cai Liang-liang Zheng Li-mou Zeng Chi-meng |
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| Institution: | WAN Wei-wei1,ZHANG Huan-jing1,YE Hui-ming1,WAN Li-qin2,CAI Liang-liang1,ZHENG Li-mou1,TZENG Chi-meng1 1Center of Translational Medicine Research,Institute For Biomedical Research,Xiamen University,Xiamen 361005,2The Third of Department of General Surgery,the First Affiliated Hospital of Nanchang University,Nanchang 330006,China |
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| Abstract: | AIM: To synthesize the 25-hydroxyvitamin D3 artificial complete antigen and to prepare the specific antibody against 25-hydroxyvitamin D3.METHODS: The active group carboxyl was introduced into 25-hydroxyvitamin D3 and formed 25-hydroxyvitamin D3-hemisuccinate which possessed the structure of the hapten by chemical modification.The EDC method was applied to conjugate 25-hydroxyvitamin D3-hemisuccinate to bovine serum albumin as an artificial immunogen.The coating antigen 25-hydroxyvitamin D3-hemisuccinate-OV... |
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| Keywords: | 25-hydroxyvitamin D3 hapten-[C-3]-carrier conjugates artificial complete antigen |
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