过氧化氢酶-氯霉素偶联物的合成及其结合比的测定

利用混合酸酐法合成了氯霉素琥珀酸酯衍生物．再用碳二亚胺将该衍生物与过氧化氢酶偶联,用紫外分光光度法测定已纯化的偶联物中氯霉素与过氧化氢酶的结合比。结果,偶联物中氯霉素与过氧化氢酶的结合比达到30．67:1,偶联物中过氧化氢酶的活性损失约30％。结果表明,该偶联物可作为竞争性ELISA法或免疫传感器法测试氯霉素时所需的酶标抗原。

Synthesis of catalase-chloramphenicol conjugation and ultraviolet determination of molar incorporation ratio of chloramphenicol into catalase

A succinyl derivation of chloramphenicol (HS-CAP) was prepared using the method of mixed anhydride reaction and the derivation was conjugated with catalase (CAT) using dual functional reagent l-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDO for the determination of residue of chloramphenicol in animal products by establishing the method of enzyme linked assay or immunosen-sor. The molar incorporation ratio of chloramphenicol into catalase in the purified conjugation product was determined by ultraviolet spectrophotometry. The results showed that the molar incorporation ratio of HS-CAP into CAT reached 30. 67 : 1 and the lose of CAT activity was only 30% or so. Thus this conjugation can be used as the enzyme-labeled antigen for the determination of chloramphenicol by ELISA or immu-nosensor.