多西环素人工抗原的合成与鉴定

Synthesis and Identification of Antigens of Doxycycline

关键词:  多西环素  半抗原  完全抗原  合成  鉴定

Key words:  doxycycline  hapten  complete antigen  synthesis  identification

摘要:

摘要：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ELISA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39.79-53.13 ug/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。

Abstract:

Abstract：To doxycycline (doxycycline, DOX) for chemical modification such as the introduction of carboxyl or amino-active group, and then with bovine serum albumin (BSA) and ovalbumin (OVA) 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 ug/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.