title: Optimization of condition for conjugation of enrofloxacin to enzymes in chemiluminescence enzyme immunoassay.
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.
authors.affiliation: College of Public Health, Zhengzhou University, Zhengzhou 450001, PR China.
authors.forename: Songcheng
authors.initials: S
authors.lastname: Yu
authors.affiliation: College of Public Health, Zhengzhou University, Zhengzhou 450001, PR China.
authors.forename: Fei
authors.initials: F
authors.lastname: Yu
authors.affiliation: College of Public Health, Zhengzhou University, Zhengzhou 450001, PR China.
authors.forename: Hongquan
authors.initials: H
authors.lastname: Zhang
authors.affiliation: College of Public Health, Zhengzhou University, Zhengzhou 450001, PR China.
authors.forename: Lingbo
authors.initials: L
authors.lastname: Qu
authors.affiliation: College of Public Health, Zhengzhou University, Zhengzhou 450001, PR China. Electronic address: wuyongjun@zzu.edu.cn.
authors.forename: Yongjun
authors.initials: Y
authors.lastname: Wu
mesh_terms: D000469:Alkaline Phosphatase / Q000737:chemistry*; D000911:Antibodies, Monoclonal / Q000737:chemistry*; D000077422:Enrofloxacin; D024841:Fluoroquinolones / Q000737:chemistry*; D006735:Horseradish Peroxidase / Q000737:chemistry; D007124:Immunoenzyme Techniques; D008163:Luminescent Measurements / Q000379:methods*
chemical_list: D000911:Antibodies, Monoclonal; D024841:Fluoroquinolones; D000077422:Enrofloxacin; D006735:Horseradish Peroxidase; D000469:Alkaline Phosphatase
keywords: Chemiluminescence enzyme immunoassay; Conjugation; Enrofloxacin; Label enzyme