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投稿时间：2015-11-29
投稿时间：2015-11-29
中文摘要: 基于抗沙丁胺醇（SAL）的单克隆抗体，初步建立了检测 SAL 的高灵敏度的时间分辨直接竞争免疫分析法（CDTRFIA）。采用辛酸-硫酸铵纯化抗 SAL 杂交瘤细胞株腹水单克隆抗体，用稀土离子 Eu3+偶联物进行标记，制备铕标抗 体； 通过合成 SAL-SUC 半抗原并与卵清蛋白偶联获得 SAL-OVA 包被抗原： 采用直接竞争的方式， 游离 SAL 和固相 SAL-OVA 包被原共同竞争有限的铕标 SAL 单抗，初步建立 SAL 时间分辨免疫分析方法。研究发现，优化的 TRFIA 半抑 制量 IC50 为 1.6 ng/mL，检测范围为 0.39 ng/mL~12.77 ng/mL，最低检测限为 0.136 ng/mL。通过对其他常用 β-兴奋剂进行 交叉反应分析，研究发现建立的 TRFIA 方法特异选择性高，与克伦特罗的交叉率较小（2.29 %），而与莱克多巴胺没有 交叉。
中文关键词: 沙丁胺醇 铕标单克隆抗体 时间分辨荧光免疫分析
Abstract:In order to provide a rapid and selectivity method for the determination of salbutamol（SAL）， an di－ rect competitive time-resolved fluorescence immunoassay（TRFIA）was developed. Anti-SAL monoclonal anti－ body was gained from hybridomas 4E4 produced before in our lab and then conjugated to Eu3+ to prepare Eu3+ -labeled antibody as a tracer in TRFIA. SAL-OVA was coated by physical adsorption onto the microtitre plate， SAL or sample with SAL as a competitor. Both of them were incubated with limited anti-SAL Eu3+ labeled an－ tibody. And coating concentration， dilution of Eu3+ -labeled antibody， competitive time and reaction model of SAL-TRFIA was optimized to obtain the better sensitivity. The method of SAL-TRFIA had an IC50 of 1.6 ng/mL， which could detect SAL in the range from 0.39 ng/mL to 12.77 ng/mL ，the detection limit was 0.136 ng/mL. The cross reactivity of the SAL-TRFIA with ractopamine was negligible， while that with clenbuterol was 2.29 %.
文章编号：201607038 中图分类号： 文献标志码：A
基金项目:
| Author Name | Affiliation |
| HOU Wen-hui | Ulanqab Vocational College， Wulanchabu 012000， Inner Mongolia， China |
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