Development of polyclonal antibody against clenbuterol for immunoassay application

• Main Authors: A. Talib, Nurul Ain, Salam, Faridah, Sulaiman, Yusran
• Format: Article
• Language: English
• Published: MDPI 2018
• Online Access: http://psasir.upm.edu.my/id/eprint/72268/1/Development%20of%20polyclonal%20antibody.pdf; http://psasir.upm.edu.my/id/eprint/72268/; https://www.mdpi.com/1420-3049/23/4/789
• Institution: Universiti Putra Malaysia

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.