Development of polyclonal antibody against clenbuterol for immunoassay application
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...
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| Main Authors: | , , |
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| Format: | Article |
| Language: | English |
| Published: |
MDPI
2018
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| 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 |
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| Institution: | Universiti Putra Malaysia |
| Language: | English |
| Summary: | 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. |
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