克伦特罗单克隆抗体的纯化及其生物学特性的研究

目的：将自制克伦特罗（CL）单克隆抗体纯化并研究其生物学特性，进行性质鉴定并建立检测标准曲线。方法：用ELISA法测定克伦特罗单克隆抗体的亲和常数和抗体活性，ELJSA测定单克隆抗体与BSA的交叉反应及与几种结构和功能类似物的交叉反应，然后采用间接竞争ELISA方法建立检测标准曲线。将制备的含克伦特罗单克隆抗体的小鼠腹水用盐析法和免疫亲和柱层析法进行抗体纯化。结果：经ELISA法测定，单克隆抗体亲和常数为2．90×10mmol／L，抗体效价最高达10^6。单克隆抗体对BSA无反应，对几种结构和功能类似物的交叉反应率均小于0．005％。建立的标准曲线R2=0．9812，最低检测限为1．0ng／ml。结论：建立了间接竞争ELISA检测cL的标准曲线。自制的克伦特罗单克隆抗体亲和力好，特异性高。为以后实际样品的检测及制备CL免疫检测试纸条和试剂盒奠定了基础。

Purification of monoclonal antibody to clenbuterol and its biology identity

Objective： To identify the self-preparation monoclonal antibody which target to clenbuterol, and set up the standard curve to clenbuterol （CL） detection. Methods： The affinity constants and activity of the monoclonal antibody which target to CL were determined by ELISA. ELISA was also used to confirm whether the monoclonal antibody had any across-reaction with BSA and CL analogues. The rat ascites which contains the monoclonal antibody target to CL was purified by （NH4）2S04 salt-out method and further by aff＇mity colunm. At last, the CL detection standard curve which based on indirect competition EI.ISA was established. Results： The ELISA experiment showed that the antibody fiter was 106 and the monoclonal antibody affinity constants was 2.90 ×1010 L/mol. The result of the indirect competition ELISA confirmed that the monoclonal antibody had no cross-reaction with BSA and a few kind of CL analogue. CL detection standard curve based on indirect compe- tition ELISA was established, which R2 was 0.9812, and the lowest detectable limit was 1.0 ng/ml. Conclusion： The standard curve based on indirectly competitioning ELISA was established. The self-preparation monoclonal antibody which target to CL has high affinity and high specific to CL, which had established the foundation to the advanced develot~nent of the CL immune test paper and CL ELISA kit.