Development of Polyclonal Antibody against Clenbuterol for Immunoassay Application

Molecules

Faridah Salam¹  Yusran Sulaiman²  Nurul Ain A. Talib² 

[1] Biodiagnostic-Biosensor Programme, Biotechnology and Nanotechnology Research Centre, Malaysian Agricultural Research and Development Institute, 43400 Serdang, Malaysia;Functional Device Laboratory, Institute of Advance Technology, Universiti Putra Malaysia, 43400 Serdang, Malaysia;

关键词: polyclonal antibody;    clenbuterol;    β-agonist;    antibody titer;    sodium dodecyl sulfate-polyacrylamide gel electrophoresis;    ELISA;   

DOI  :  10.3390/molecules23040789

来源: DOAJ

【 摘 要 】

Development of an immunoassay for clenbuterol (CLB) detection required an anti-CLB antibody as an important bioreceptor. In this study, we report our work on production and purification of a rabbit-derived polyclonal anti-CLB antibody. The antibody was then purified by nProtein A Sepharose affinity column and the antibody purity was confirmed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis. The activities of purified antibody were evaluated based on high antibody titer determined from enzyme-linked immunosorbent assay (ELISA). The sensitivity and selectivity of this antibody was evaluated and exhibits negligible cross-reactivity to antibiotics other than β-agonist families. Evaluation of the antibody as bioreceptor in immunoassay was performed using direct competitive ELISA and exhibited linear calibration plot (R2 = 0.9484). The antibody was used to detect the content of CLB in spiked milk samples and the recovery of more than 92% indicating significant performance as bioreceptor for the development of a rapid and simple immunoassay.

【 授权许可】

Unknown