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过氧化氢酶-氯霉素偶联物的合成及其结合比的测定
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柴春彦，徐明刚，刘国艳
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（上海交通大学 农业与生物学院 动物科学系，上海201101）
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摘要：
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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。 |
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关键词：
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氯霉素；过氧化氢酶；结合比；酶标抗原 |
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中图分类号：S 859.796 文献标识码：
A 文章编号：1673-4696(2006)5-0406-05 |
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Synthesis of catalase-chloramphenicol conjugation and
ultraviolet determination of molar incorporation ratio of
chloramphenicol into catalase |
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CHAI Chun-yan，XU Ming-gang，LIU Guo-yan |
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(Department of Animal Science, School of Agriculture and Biology, Shanghai Jiaotong University,
Shanghai 201101, China) |
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Abstract: 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 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDC) for the determination of residue of chloramphenicol in animal products by establishing the method of enzyme linked assay or immunosensor. 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 3067∶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 immunosensor. |
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Key words: chloramphenicol; catalase; incorporation ratio;enzyme-labeled antigen |
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