Abstract：
Florfenicol (FF), with its broad-spectrum antibacterial activity, is frequently abused in the livestock and poultry industries and has aroused the growing public concern. Owing to structural similarities and varying maximum residue limits between florfenicol and other chloramphenicol (CAP)-type antibiotics, including thiamphenicol (TAP) and chloramphenicol (CAP), there is an urgent need for a rapid and effective immunoassay method to distinguish them, in order to minimize the risk of false positives. Fortunately, a highly specific monoclonal antibody (mAb), named as SF11, has been developed using hybridoma technology. Molecular simulations have revealed that the mAb SF11\'s specificity in recognizing florfenicol stems from the π-π stacking interaction between florfenicol and the mAb SF11 binding pocket. Using this highly specific mAb, a sensitive time-resolved fluorescence immunochromatographic assay (TRFICA) strip for rapid florfenicol detection has been developed. Under optimal conditions, this TRFICA demonstrated good analytical performance for the detection of florfenicol in milk and eggs samples, with the half-maximal inhibition concentration (IC50) values of 1.89 and 2.86 ng mL-1, the limit of detection (LOD) of 0.23 and 0.48 ng mL-1, the cut-off values of 62.50 and 31.25 ng mL-1, and the testing time of approximately thirteen minutes. Spiked recoveries in the milk and eggs samples ranged from 104.7 % to 112.3 % and 95.3 % to 116.4 %, respectively, with no obvious cross-reactions with the other analogues observed. The TRFICA results correlated well with those of high-performance liquid chromatography tandem mass spectrometry (HPLC-MS/MS) for real samples, indicating that the developed TRFICA method was sensitive, accurate and adapted for the rapid determination of florfenicol in milk and egg samples.
摘要：
氟苯尼考(FF),具有广谱抗菌活性,在畜禽行业中经常被滥用，并引起了公众的日益关注。由于氟苯尼考和其他氯霉素（CAP）型抗生素之间的结构相似性和不同的最大残留限值，包括甲砜霉素（TAP）和氯霉素（CAP），迫切需要一种快速有效的免疫分析方法来区分它们，以尽量减少误报的风险。幸运的是,一种高度特异性的单克隆抗体（mAb），命名为SF11，已开发使用杂交瘤技术。分子模拟表明，mAbSF11识别氟苯尼考的特异性源于氟苯尼考与mAbSF11结合袋之间的π-π堆叠相互作用。使用这种高度特异性的单克隆抗体，已经开发了用于快速检测氟苯尼考的灵敏的时间分辨荧光免疫色谱测定（TRFICA）条。在最优条件下,该TRFICA在检测牛奶和鸡蛋样品中氟苯尼考方面表现出良好的分析性能，半最大抑制浓度（IC50）值为1.89和2.86ngmL-1，检测限（LOD）为0.23和0.48ngmL-1，临界值为62.50和31.25ngmL-1，测试时间约为13分钟。牛奶和鸡蛋样品的加标回收率为104.7%至112.3%，95.3%至116.4%，分别,与其他类似物没有明显的交叉反应。TRFICA结果与实际样品的高效液相色谱串联质谱(HPLC-MS/MS)表明所开发的TRFICA方法是灵敏的，准确，适用于牛奶和鸡蛋样品中氟苯尼考的快速测定。
