西马特罗人工抗原的合成及鼠源多克隆抗血清的制备

Synthesis of Cimaterol Artificial Antigen and Development of Its Polyclonal Antiserum

拟合成并鉴定西马特罗(Cimaterol,CIM)人工抗原,通过动物免疫生产亲和力高、特异性好的鼠源CIM多克隆抗血清。通过重氮化方法将西马特罗偶联于载体蛋白BSA和OVA上,分别合成免疫抗原BSA-CIM和包被抗原OVA-CIM,并采用紫外分光光度法和SDS-PAGE进行了鉴定。结果表明,半抗原CIM和载体蛋白偶联成功。动物免疫试验结果显示,6只小鼠效价均达1×10-3以上,且4号小鼠多抗血清抗CIM的IC50为86.9ng/mL,表明成功获得了高效价、特异性好、亲和力高的的鼠源抗CIM多抗血清。

The goal of this study was to synthesize artificial antigen of cimaterol and obtain its mice polyclonal antiserum. Artificial antigen BSA-CIM and OVA-CIM were synthesized using diazotization by linking carrier proteins BSA and OVA to CIM. The antigen BSA-CIM and OVA-CIM was identified by ultraviolet scanning,SDS-PAGE. The results showed that the hapten CIM was successfully linked to carrier proteins. The animal immunization results showed that six BALB /c mice indirect ELISA titer against CIM were above 1 × 10^ -3 and the IC50 of No. 4 mice was 86. 9 ng /mL and the high-titer,sensitivity CIM polyclonal antiserum has been generated. The artificial antigen of CIM and its mice polyclonal antiserum laid a foundation base for product the monoclonal antibodies and rapid test reagent against CIM.