ABSTRACT Polyclonal antibodies were obtained by immunizing rabbits with polymyxin B (PMB) conjugated to bovine serum albumin. Based on the heterologous coating conjugate of PMB with periodate-oxidized gelatin and the resulting antibodies, an indirect competitive enzyme-linked immunosorbent assay (ELISA) was developed to detect residual colistin (COL), a structural analogue of PMB in livestock products. The developed analysis was characterized by close specificity towards PMB (100%) and COL (88%), sensitivity (IC50 = 5.7 ng/mL), dynamic range (IC20-IC80 = 1.0–80 ng/mL) and the detection limit of COL (IC10 = 0.4 ng/mL). It was possible to quantify the antibiotic in milk and eggs without special and labour-intensive sample pretreatment using a simple 10- and 50-fold dilution of the corresponding samples. The developed analysis system made it possible to record the maximum residue limit of COL in dairy products (50 μg/kg) and eggs (300 μg/kg) with 89–104% recovery of analyte.

中文翻译：

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用于牛奶和鸡蛋分析的粘菌素间接竞争性酶联免疫分析的开发

摘要 通过使用与牛血清白蛋白偶联的多粘菌素 B (PMB) 对兔子进行免疫接种来获得多克隆抗体。基于 PMB 与高碘酸盐氧化明胶的异源涂层偶联物和由此产生的抗体，开发了一种间接竞争性酶联免疫吸附测定 (ELISA) 来检测残留的粘菌素 (COL)，这是一种畜产品中 PMB 的结构类似物。开发的分析的特点是对 PMB (100%) 和 COL (88%) 的密切特异性、灵敏度 (IC50 = 5.7 ng/mL)、动态范围 (IC20-IC80 = 1.0–80 ng/mL) 和检测限COL (IC10 = 0.4 ng/mL)。无需对相应样品进行 10 倍和 50 倍的简单稀释，无需进行特殊的劳动密集型样品预处理，即可对牛奶和鸡蛋中的抗生素进行定量。