米诺地尔多克隆抗体的制备及其酶联免疫检测方法的建立

Preparation of polyclonal antibody for minoxidil,development of an ELISA method for detection of minoxidil

目的：在人工合成米诺地尔抗原基础上,制备其多克隆抗体并建立相应酶联免疫检测方法。方法：采用戊二醛法合成米诺地尔人工抗原,并通过紫外扫描进行鉴定。用免疫原（Minoxidil-BSA）免疫BALB/c雌性小鼠,制备抗minoxidil的多克隆抗体,并鉴定其效价。建立检测米诺地尔的竞争ELISA法。结果：紫外扫描结果表明米诺地尔成功地偶联到载体蛋白上。免疫制备得到的多克隆抗体,其效价为1∶12 800。其酶联免疫检测曲线方程y=-0.0 823x＋0.938（R2=0.9811）,minoxidil质量浓度与吸光度在0.001-10μg/ml范围内呈良好的线性关系。结论：本研究成功制备米诺地尔人工抗原及其多克隆抗体并建立了酶联免疫检测方法,为米诺地尔免疫检测方法的实际应用奠定了基础。

Objective： On the basis of synthetic antigens on minoxidil,we prepared its polyclonal antibody and established a ELISA method to detect minoxidil. Methods： We synthesized minoxidil artificial antigen with glutaraldehyde and identified by UV scanning. We had established a detection of minoxidil competition ELISA. Results： The UV scan showed that the minoxidil successfully coupled to a carrier protein. The conjugate of minoxidil-BSA was used to immunize BALB / c mice. And the produced antiserum showed high titer of 1∶ 12 800 in the indirect ELISA. Its ELISA curve equation： y =- 0. 082 3x ＋ 0. 938（ R2= 0. 9811）,minoxidil mass concentration and the absorbance in 0. 001- 10 μg / ml showed a good linearity. Conclusion： This method has been successfully preparing minoxidil artificial antigen and polyclonal antibodies and established ELISA detection method for minoxidil,which provides a basis for the practical application of minoxidil immunoassay method.