摘要
采用琥珀酸酐法合成喹乙醇半琥珀酸酯(OLA-HS),以质谱法对其进行鉴定;活化酯法合成全抗原OLA-BSA、OLA-OVA,以紫外(UV)和凝胶电泳(SDS-PAGE)鉴定之;用OLA-BSA免疫BALB/c小鼠,间接ELISA测定多抗血清效价,阻断ELISA鉴定其敏感性、特异性。紫外扫描测得OLA-BSA、OLA-OVA中OLA-HS和BSA、OVA的分子结合比分别为17.1∶1和12.4∶1;3号小鼠抗血清的效价最高,对OLA的IC50为58.923 ng/mL,与卡巴氧的交叉反应率为1.841%,与其他药物无交叉反应。通过系列ELISA鉴定,获得高价、敏感、特异的多克隆抗体(pAb),为OLA残留免疫学检测方法的建立奠定基础。
The derivative of olaquindox,called as OLA-SH,was synthesized by succinic anhydride method and identified by mass spectrometry（MS）.Two complete antigens of olaquindox（OLA-BSA,OLA-OVA） were prepared by active ester method（AE）.The characterization of hapten-protein conjugates were examined by two methods,ultraviolet spectrophotometry（UV）,and sodium dodecyl sulfate polyacrylamide gel electrophoresis（SDS-PAGE）,respectively.BALB/C mice were immunized with OLA-BSA,the titre of polyclonal antibody（pAb）was detected by indirect ELISA,the sensitivity and specificity of pAb was identified by blocking ELISA.According to UV scanning,the conjugation ratio of OLA-HS to BSA and OVA was about 17.1∶1 and 12.4∶1.The titer of pAb for No.3 mouse was the highest,its IC50 was 58.923 ng/mL,cross reaction rate to carbadox was 1.841%,and no cross reaction to other antibiotics.The high-titer,sensitive and specific anti-OLA pAb have been produced in sera of ELISA,which made it possible to establish immunoassay of OLA residues.
出处
《西北农业学报》
CAS
CSCD
北大核心
2010年第7期1-6,共6页
Acta Agriculturae Boreali-occidentalis Sinica
基金
国家“十一五”科技支撑计划项目(2006BAK02A21)
关键词
喹乙醇
人工抗原
合成
鉴定
Olaquindox
Artificial antigen
Synthesis
Identification
