التفاصيل البيبلوغرافية
| العنوان: |
25-羟基维生素D_3人工完全抗原的合成与鉴定 ; Synthesis and identification of artificial complete antigen 25-hydroxyvitamin D_3 |
| المؤلفون: |
万巍巍, 张换敬, 叶辉铭, 万丽琴, 蔡良良, 郑立谋, 曾骥孟 |
| سنة النشر: |
2011 |
| المجموعة: |
Xiamen University Institutional Repository |
| مصطلحات موضوعية: |
25-羟基维生素D3, 半抗原-[C-3]-载体蛋白偶联, 人工完全抗原, 25-hydroxyvitamin D3, hapten-[C-3]-carrier conjugates, artificial complete antigen |
| الوصف: |
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6.25x10-4)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。 ; 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-OVA was obtained in the same way.Ultraviolet,SDS-PAGE and MALDI-Tof were used to identify 25-hydroxyvitamin D3-hemisuccinate-BSA.RESULTS: BALB/c mice were immunized with 25-hydroxyvitamin D3-hemisuccinate-BSA to generate the polyclonal antibody of the 25-hydroxyvitamin D3 worth high titer and the immunogen,25-hydroxyvitamin D3-hemisuccinate-BSA,was successfully prepared with coupling ratio(12±0.16)∶1(N=3) coupling.CONCLUSION: The high titer and good sensitivity of anti-25-hydroxyvitamin D3 antibody are produced in sera immunized BALB/c mice,which made it possible to develop a clinical diagnostics for illness. |
| نوع الوثيقة: |
article in journal/newspaper |
| اللغة: |
Chinese |
| تدمد: |
1007-8738 |
| Relation: |
细胞与分子免疫学杂志,2011,(11):13-15+19; XBFM201111004; http://dspace.xmu.edu.cn/handle/2288/119670 |
| الاتاحة: |
http://dspace.xmu.edu.cn/handle/2288/119670 |
| رقم الانضمام: |
edsbas.BF5170E4 |
| قاعدة البيانات: |
BASE |