Production of the monoclonal antibody against tylosin for immunoassay of macrolide antibiotics in milk

Abstract

The objective of the present study was to develop an immunoassay for multi-determination of the residues of four macrolide antibiotics in milk. Tylosin was derivatized with 6-aminohexanoic acid to synthesize a hapten that was used to produce the monoclonal antibody. The obtained monoclonal antibody simultaneously recognized tylosin, acetylisovaleryltylosin, tilmicosin and desmycosin with crossreactivities of 100%, 91%, 49% and 76%, respectively. Then an indirect competitive immunoassay was developed to determine the four analytes in milk simultaneously. The limits of detection for the four analytes were in the range of 5.5–11.7 ng/mL depending on the compound. The recoveries of the four analytes from fortified blank milk were in the range of 78%–96% with coefficients of variation lower than 11.4%. The developed immunoassay could be used as a practical tool for rapid screening the residues of the four anlytes in milk.

Keywords:

• Macrolide antibiotics
• tylosin
• monoclonal antibody
• immunoassay
• milk

Acknowledgment

The authors are grateful for the financial support of Hebei Scientific and Technological Project (11221001D).

The two authors J. K. Zhang and J. X. Liu contributed equally to this work.