| 大田软海绵酸人工抗原的合成及其多克隆抗体的制备 |
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| 引用本文： | 王丽,桑亚新,周群标,王向红.大田软海绵酸人工抗原的合成及其多克隆抗体的制备[J].水产学报,2011,35(7):1001-1007. |
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| 作者姓名： | 王丽 桑亚新 周群标 王向红 |
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| 作者单位： | 河北农业大学食品科技学院,河北省农产品加工工程技术研究中心,河北,保定,071001 |
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| 基金项目： | 国家海洋局公益性行业科研专项(200805046) |
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| 摘 要： | 采用活化酯法将大田软海绵酸(OA)与匙孔血蓝蛋白(KLH)偶联制备免疫抗原(OA-KLH),用还原聚丙烯酰胺凝胶电泳法和红外光谱法对人工抗原的偶联效果进行分析,通过免疫兔子制备抗体,所得抗血清经Protein A凝胶层析柱纯化处理后,用紫外全波长扫描和间接竞争ELISA法验证纯化效果。结果表明,免疫抗原偶联成功并获得了高亲和力的多克隆抗血清,抗血清纯化后浓度为2.16 mg/mL,间接竞争ELISA测定其滴度为12 800、IC15为3.41 ng/mL,为建立快速、经济的检测方法打下了基础。
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| 关 键 词： | 大田软海绵酸 人工抗原 红外表征 抗体纯化 |
| 收稿时间： | 2010/12/16 0:00:00 |
| 修稿时间： | 2011/5/11 0:00:00 |
Synthesis of artificial antigen and preparation of polyclonal antibody against okadaic acid |
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| WANG Li,SANG Ya-xin,ZHOU Qun-biao and WANG Xiang-hong.Synthesis of artificial antigen and preparation of polyclonal antibody against okadaic acid[J].Journal of Fisheries of China,2011,35(7):1001-1007. |
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| Authors: | WANG Li SANG Ya-xin ZHOU Qun-biao and WANG Xiang-hong |
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| Institution: | Agriculture University of Hebei,Agriculture University of Hebei,Agriculture University of Hebei,Agriculture University of Hebei |
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| Abstract: | Okadaic acid(OA)is one kind of lipophilic marine biotoxins,and main pathogenic factor of diarrheic shellfish poisoning(DSP).In this paper,the artificial antigen was made by coupling hapten of OA with keyhole limpet hemocyanin(KLH)using active ester method,and the artificial antigen was tested by SDS-polyacrylamide gel electrophoresis and IR spectrum.After being obtained from rabbits,the antiserum was purified by Chromatography on Protein A Sepharose CL-4B gel column,and the purification efficiency was verified by ultraviolet scanning and ciELISA.The results showed the artificial antigen was prepared successfully and lots of interfering substances were removed.After being purified,the concentration of antiserum was 2.16 mg/mL,it had high titer(12 800)and sensitivity(the value of IC15 was 3.41 ng/mL),which was tested by ciELISA.The success of obtaining polyclonal antibody against OA with high titer and affinity provides the solid foundation which is practical and theoretical for developing rapid and economical detections. |
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| Keywords: | okadaic acid(OA) antigen synthesis IR characterization antibody purification |
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