Novel hapten synthesis for antibody production and development of an enzyme-linked immunosorbent assay for determination of furaltadone metabolite 3-amino-5-morpholinomethyl-2-oxazolidinone (AMOZ)
Novel hapten synthesis for antibody production and development of an enzyme-linked immunosorbent assay for determination of furaltadone metabolite 3-amino-5-morpholinomethyl-2-oxazolidinone (AMOZ)
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用于抗体生产的新型半抗原合成以及用于测定呋喃他酮代谢物 3-氨基-5-吗啉代甲基-2-恶唑烷酮 (AMOZ) 的酶联免疫吸附测定法的开发
DOI：
10.1016/j.talanta.2012.10.059
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发表时间：
2013-01-15
期刊：
影响因子：
6.1
通讯作者：
Jiang, Yue-Ming
中科院分区：
文献类型：
--
作者：
Xu, Zhen-Lin;Shen, Yu-Dong;Jiang, Yue-Ming
开发了一种异源竞争性间接酶联免疫吸附测定 (ciELISA)，用于测定呋喃他酮代谢物 3-氨基-5-吗啉代甲基-2-恶唑烷酮 (AMOZ)。 AMOZ用2-(4-甲酰基苯氧基)乙酸或2-(3-甲酰基苯氧基)乙酸衍生化以获得两种新型免疫半抗原。将这些半抗原产生针对 AMOZ 硝基苯基衍生物 (NPAMOZ) 的特异性多克隆抗体的能力与传统免疫半抗原（用 3-羧基苯甲醛或 4-羧基苯甲醛衍生的 AMOZ）进行比较。结果表明，新型免疫半抗原能够产生抗体，与现有基于抗体的 ELISA 相比，ciELISA 对 NPAMOZ 的灵敏度几乎提高了两倍。灵敏度的差异可以通过 NPAMOZ 和半抗原最低能量构象的分子模型来解释。合成了另一种新型半抗原，用 2-氧代乙酸衍生的 AMOZ，并将其用作异源涂层半抗原。结果表明，这种仅使用目标分子的部分结构作为涂层半抗原的策略能够使灵敏度提高两到三倍。这项研究为免疫分析的开发提供了一种现代方法，提高了硝基呋喃抗生素代谢物的灵敏度。 (C) 2012 Elsevier B.V. 保留所有权利。
A heterologous competitive indirect enzyme-linked immunosorbent assay (ciELISA) for the determination of the furaltadone metabolite 3-amino-5-morpholinomethyl-2-oxazolidinone (AMOZ) was developed. AMOZ was derivatised with 2-(4-formylphenoxy) acetic acid or 2-(3-formylphenoxy) acetic acid to obtain two novel immunizing haptens. The ability of these haptens in producing specific polyclonal antibodies against the nitrophenyl derivative of AMOZ (NPAMOZ) was compared with that of traditional immunizing haptens (derivatised AMOZ with 3-carboxybenzaldehyle or 4-carboxybenzaldehyle). The results indicated that the novel immunizing haptens were able to produce antibodies with almost a two-fold improvement in sensitivity of the ciELISA for NPAMOZ in comparison with the existing antibody based ELISAs. The differences in sensitivity were explained by the molecular modeling of the lowest energy conformations of NPAMOZ and the haptens. Another novel hapten, derivatised AMOZ with 2-oxoacetic acid, was synthesized and used as a heterologous coating hapten. The results showed that this strategy of using only a partial structure of the target molecule as the coating hapten was able to obtain a two to three-fold improvement in sensitivity. This study provided a modern approach for the development of an immunoassay with improved sensitivity for the metabolites of nitrofuran antibiotics. (C) 2012 Elsevier B.V. All rights reserved.