化学发光酶免疫分析中恩诺沙星与酶偶联条件的优化。

Optimization of condition for conjugation of enrofloxacin to enzymes in chemiluminescence enzyme immunoassay.

作者信息

Yu Songcheng, Yu Fei, Zhang Hongquan, Qu Lingbo, Wu Yongjun

机构信息

College of Public Health, Zhengzhou University, Zhengzhou 450001, PR China.

出版信息

Spectrochim Acta A Mol Biomol Spectrosc. 2014 Jun 5;127:47-51. doi: 10.1016/j.saa.2014.02.019. Epub 2014 Feb 22.

DOI:10.1016/j.saa.2014.02.019

PMID:24632155

Abstract

In this study, in order to find out a proper method for conjugation of enrofloxacin to label enzymes, two methods were compared and carbodiimide condensation was proved to be better. The results showed that the binding ratio of enrofloxacin and alkaline phosphatase (ALP) was 8:1 and that of enrofloxacin and horseradish peroxidase (HRP) was 5:1. This indicated that conjugate synthesized by carbodiimide condensation was fit for chemiluminescence enzyme immunoassay (CLEIA). Furthermore, data revealed that dialysis time was an important parameter for conjugation and 6days was best. Buffer to dilute conjugate had little effect on CLEIA. The storage condition for conjugates was also studied and it was shown that the conjugate was stable at 4°C with no additive up to 30days. These data were valuable for establishing CLEIA to quantify enrofloxacin.

摘要

在本研究中，为找出恩诺沙星与标记酶偶联的合适方法，对两种方法进行了比较，结果证明碳二亚胺缩合法更好。结果表明，恩诺沙星与碱性磷酸酶（ALP）的结合比为8:1，恩诺沙星与辣根过氧化物酶（HRP）的结合比为5:1。这表明通过碳二亚胺缩合合成的偶联物适用于化学发光酶免疫分析（CLEIA）。此外，数据显示透析时间是偶联的一个重要参数，6天为最佳。稀释偶联物的缓冲液对CLEIA影响不大。还研究了偶联物的储存条件，结果表明，在不加添加剂的情况下，偶联物在4℃可稳定保存30天。这些数据对于建立定量恩诺沙星的CLEIA具有重要价值。