School of Biosystems Engineering and Food Science, Zhejiang University, Hangzhou, People's Republic of China.
J Sci Food Agric. 2013 Feb;93(3):667-73. doi: 10.1002/jsfa.5945. Epub 2012 Nov 26.
Monoclonal antibodies generally obtained through the classic mouse hybridoma system were requisite for the establishment of various immunoassays. In this study, a new rabbit monoclonal antibody (RabMAb) against sulfonamides (SAs) was first produced via hybridoma technique in rabbit. The related enzyme-linked immunosorbent assay (ELISA) was then developed and applied to real sample analysis.
A sensitive competitive indirect ELISA method based on a novel RabMAb for rapid detection of sulfonamides was first established. The obtained half-maximum inhibition concentration (IC(50)) values for four SAs were all below 10 ng mL(-1) , with 0.68 ng mL(-1) sulfathiazole (STZ), 1.11 ng mL(-1) sulfadiazine (SD), 1.15 ng mL(-1) sulfapyridine (SP) and 5.27 ng mL(-1) sulfamethoxazole (SMX). Desirable recoveries when detecting different spiked swine urine and milk samples were achieved ranging from 92.6% to 104.3% and from 61.1% to 81.6%, respectively.
The proposed immunoassay with the newly developed RabMAb is capable of detection of four SAs (STZ, SD, SP and SMX) with proven satisfactory performance and is applicable for routine large-scale analysis in practical uses.
通过经典的小鼠杂交瘤系统获得的单克隆抗体通常是建立各种免疫分析所必需的。在这项研究中，首次通过杂交瘤技术在兔子中产生了一种针对磺胺类药物（SAs）的新型兔单克隆抗体（RabMAb）。然后开发了相关的酶联免疫吸附测定（ELISA），并将其应用于实际样品分析。
首次建立了一种基于新型 RabMAb 的快速检测磺胺类药物的灵敏竞争间接 ELISA 方法。四种磺胺类药物的半最大抑制浓度（IC50）值均低于 10ng/mL，其中磺胺噻唑（STZ）为 0.68ng/mL，磺胺嘧啶（SD）为 1.11ng/mL，磺胺吡啶（SP）为 1.15ng/mL，磺胺甲恶唑（SMX）为 5.27ng/mL。在检测不同浓度添加的猪尿液和牛奶样品时，得到了令人满意的回收率，范围分别为 92.6%至 104.3%和 61.1%至 81.6%。
该免疫分析方法具有良好的性能，能够检测四种磺胺类药物（STZ、SD、SP 和 SMX），适用于实际应用中的常规大规模分析。
