多西环素人工抗原的合成与鉴定

将多西环素（doxycycline，DOX）进行化学修饰引入羧基或氨基等活性基团，然后与牛血清白蛋白（BSA）和卵清蛋白（OVA）偶联合成人工免疫原DOX—PABA—BSA、DOX—BSA和包被原DOX—PABA—OVA、DOX～OVA，并用紫外吸收（UV）、凝胶电泳（SDS—PAGE）和EUSA方法对人工抗原进行鉴定；将合成的人工抗原DOX—PABA—BSA、DOX—BSA分别免疫BALB／C小鼠，用间接ELISA方法测定多抗（pAb）效价，用竞争ELISA方法鉴定其敏感性，用交叉反应试验鉴定其特异性。结果表明，二个免疫组6只小鼠血清抗体效价均在1：6400以上，DOXpAb对DOX的50％抑制浓度（IC50）在39．79～53．13μg/L，抗血清与四环素类药物交叉反应很低。本实验为建立多西环素ELISA残留免疫学检测方法和并制备多西环素试剂盒奠定了基础。

Synthesis and Identification of Antigens of Doxycycline

Doxycycline（DOX） was modified with carboxyl or amino -active group, and then with bovine serum albumin（BSA） and ovalbumin（OVA）which were coupled synthesis of artificial immunogen DOX- PABA- BSA, DOX - BSA and coated antigen DOX - PABA - OVA, DOX - OVA. UV absorption （ UV）, sodium dodecyl sulfate polyacrylamide gel electrophoresis （SDS -PAGE） and ELISA were identified on the artificial antigen. The synthesis of artificial antigen DOX -PABA -BSA, DOX -BSA were immunized BALB/C mice. Polyclonal antibody titer were measured by indirect ELISA, using competitive ELISA sensitivity was identified, using cross - reaction tests identify their specificity. The results showed that two immunization groups of six mice with anti - serum titer higher than 1 ： 6400 by indirect ELISA, 50% inhibitive concentration （ IC50） was 39.79 - 53.13 μg/L and low levels of cross - reactivity to Tetracyclines were observed. In this study, In this study, the Pab （anti -DOX antiserum） can be used to develop a ELISA kit to detect DOX residues in animal edible tissues.