克伦特罗多克隆抗体的研制及其在免疫分析中的应用。
Development of Polyclonal Antibody against Clenbuterol for Immunoassay Application.
机构信息
Functional Device Laboratory, Institute of Advance Technology, Universiti Putra Malaysia, 43400 Serdang, Malaysia.
Department of Chemistry, Faculty of Science, Universiti Putra Malaysia, 43400 Serdang, Malaysia.
出版信息
Molecules. 2018 Mar 29;23(4):789. doi: 10.3390/molecules23040789.
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.
为了开发用于检测克伦特罗（CLB）的免疫分析方法，需要将克伦特罗抗体作为重要的生物受体。在本研究中，我们报告了兔源多克隆抗 CLB 抗体的生产和纯化工作。然后，通过 nProtein A Sepharose 亲和柱对抗体进行纯化，并通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）分析确认抗体的纯度。根据酶联免疫吸附测定（ELISA）确定的高抗体效价评估纯化抗体的活性。评估了该抗体作为免疫分析中的生物受体的灵敏度和选择性，并且对除β-激动剂家族以外的抗生素表现出可忽略不计的交叉反应性。使用直接竞争 ELISA 评估抗体作为生物受体的性能，并显示出线性校准曲线（R²=0.9484）。该抗体用于检测添加到牛奶样品中的 CLB 含量，回收率超过 92%，表明其作为生物受体在开发快速简单的免疫分析方面具有重要性能。
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