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软骨藻酸多克隆抗体的制备
刘元嫄1, 程金平1, 高利利1, 徐 玮1, 吴浩东1
上海交通大学环境科学与工程学院
摘要:
为建立软骨藻酸(DA)的免疫检测法, 本研究采用活泼酯法将DA 与载体蛋白KLH(BSA)偶联形成完全免疫抗原(包被抗原), 并通过紫外光谱扫描证明偶联成功与否。用完全免疫抗原DA-KLH 对新西兰白兔进行免疫, 间接ELISA 法测定血清效价。实验结果表明偶联成功, 经过19 周免疫后血清效价达到1:1600, 纯化后获得效价为1:800 的多克隆抗体, 同时也间接证明了合成的完全抗原具有免疫原性,为以后建立免疫分析法奠定了基础。
关键词:  软骨藻酸  多克隆抗体  活泼酯法
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基金项目:上海科委科技攻关项目(08DZ1206302)
Preparation of polyclonal antibodies against domoic acid
Abstract:
To develop an immunoassay of domoic acid (DA), DA was coupled to keyhole limpet hemocyanin (KLH) or bovine serum albumin (BSA) by the activation ester method. The conjugates were characterized by UV absorption. Polyclonal antibodies for domoic acid were generated from rabbits after the animals had been immunized by DA-KLH for 19 weeks. The titer of antiserum was 1:1600, detected by an indirect ELISA. After purification, the titer of antiserum became 1:800. DA-KLH was proved to be a suitable immunogen to prepare antibodies against DA.
Key words:  domoic acid  polyclonal antibody  activation ester method
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