氧氟沙星免疫亲和色谱柱的研制

Design and Study of Immunoaffinity Chromatography Column for Ofloxacin

本试验将氧氟沙星与载体蛋白偶联成人工抗原,用合成的免疫原免疫家兔,制备出高效价抗OFL血清。经纯化后研制出针对OFL特异性吸附的免疫亲和色谱柱,并对色谱柱进行了评价,偶联率为93%,动态柱容量为2204ng/mL,分别添加5mL200ng/mL、300ng/mL和400ng/mL氧氟沙星标准工作液,5mL甲醇洗脱,高效毛细管电泳检测,连续测定6次,测得其回收率为94%、92.67%、95.5%,变异系数为5.32%、3.17%、3.84%。

In this study, ofloxacin was linked respectively to cartier proteins, and artificial antigens were synthesized successfully. Using the artificial immunogens, rabbits were immunized to obtain the antiserum against ofloxacin. With purified antiserum, the immunoaffinity chromatography column was prepared for ofloxacin. After evaluation of the column, the coupling rate was determined as 93% and the dynamic capacity was 220ng/mL for ofloxacin. Using 5mL methanol as elution buffer, the recoveries were 94 %,92.6 %,95.5 %and C.V. was 5.32%, 3.17%,3.84% at levels of 200ng/mL,300ng/mL,400ng/mL in 5mL standard working liquid detected by HPCE for six times.