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

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

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.

Keywords: Chemiluminescence enzyme immunoassay; Conjugation; Enrofloxacin; Label enzyme.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alkaline Phosphatase / chemistry*
  • Antibodies, Monoclonal / chemistry*
  • Enrofloxacin
  • Fluoroquinolones / chemistry*
  • Horseradish Peroxidase / chemistry
  • Immunoenzyme Techniques
  • Luminescent Measurements / methods*

Substances

  • Antibodies, Monoclonal
  • Fluoroquinolones
  • Enrofloxacin
  • Horseradish Peroxidase
  • Alkaline Phosphatase