To avoid false-positive results in immunoassays due to cross-reactivity of antibodies with structural analogues, especially metabolites of target compounds, the preparation of highly specific antibodies is crucial. Preserving the characteristic structure of a target compound when designing a hapten is important when preparing highly specific antibodies. Here, we designed a novel hapten, 4-(((1,5-dimethyl-3-oxo-2-phenyl-2,3-dihydro-1H-pyrazol-4yl)amino)methyl)benzoic acid, named AA-BA, to improve the specificity of antibodies for detection of 4-methylaminoantipyrine (MAA), a residual marker of dipyrone, an important antipyretic-analgesic and anti-inflammatory drug. The structural features of the hapten remained almost the same as those of MAA. After experimental validation, monoclonal antibody 6A4 (mAb 6A4) was prepared with the half maximal inhibitory concentration (IC₅₀) value of 4.03 ng/mL and negligible cross-reactivity with dipyrone metabolites and other antibiotics. In addition, a specific lateral flow immunoassay (LFA) strip based on colloidal gold was developed for screening MAA with a cutoff value of 25 ng/mL in milk. The developed LFA is a useful tool for rapid and accurate detection of MAA.

中文翻译：

---

开发基于高特异性单克隆抗体的侧流免疫分析法来检测 4-甲氨基安替比林

为了避免由于抗体与结构类似物，特别是目标化合物的代谢物发生交叉反应而导致免疫测定中的假阳性结果，高度特异性的抗体的制备至关重要。在制备高度特异性抗体时，设计半抗原时保留目标化合物的特征结构非常重要。在这里，我们设计了一种新型半抗原，4-(((1,5-二甲基-3-氧代-2-苯基-2,3-二氢-1 H-吡唑-4基)氨基)甲基)苯甲酸，命名为AA- BA，提高抗体检测4-甲氨基安替比林（MAA）的特异性，MAA是安乃近的残留标志物，安乃近是一种重要的解热镇痛抗炎药物。半抗原的结构特征几乎与MAA相同。经过实验验证，制备的单克隆抗体6A4（mAb 6A4）的半数抑制浓度（IC ₅₀）值为4.03 ng/mL，与安乃近代谢物及其他抗生素的交叉反应可忽略不计。此外，还开发了一种基于胶体金的特异性侧流免疫分析 (LFA) 试纸条，用于筛选牛奶中截止值为 25 ng/mL 的 MAA。开发的 LFA 是快速准确检测 MAA 的有用工具。