State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan 430070, China.
Agricultural Bioinformatics Key Laboratory of Hubei Province, College of Informatics, Huazhong Agricultural University, Wuhan 430070, China.
Molecules. 2018 Mar 15;23(3):663. doi: 10.3390/molecules23030663.
Detection of triphenylmethane dyes (TDs), especially the widely used malachite green (MG) and crystal violet (CV), plays an important role in safety control of aquatic products. There are two chromatic forms of TDs: oxidized or reduced. Usually, only one form can be detected by reported ELISA antibodies. In this article, molecular shape superimposing and quantum mechanics calculation were employed to elucidate the differences between MG, CV, and their reduced chromatic forms (leucomalachite green, LMG and leucocrystal violet, LCV). A potential hapten was rationally designed and synthesized. Polyclonal antibodies were raised through immunizing New Zealand white rabbits and BALB/C mice. We tested the cross-reactivity ratios between the hapten and TDs. The cross-reactivity ratios were correlated with the difference in surface electrostatic potential. The determination coefficients (²) of the correlations are 0.901 and 0.813 for the rabbit and mouse antibody, respectively. According to this linear model, the significant difference in the atomic charge seemed to make it impossible to find a hapten that can produce antibodies with good cross-reactivities with both reduced and oxidized TDs.
三苯甲烷染料（TDs）的检测，特别是广泛使用的孔雀石绿（MG）和结晶紫（CV），在水产品安全控制中起着重要作用。TDs 有两种显色形式：氧化或还原。通常，报道的 ELISA 抗体只能检测到一种形式。在本文中，采用分子形状叠加和量子力学计算的方法，阐明了 MG、CV 及其还原显色形式（隐色孔雀石绿，LMG 和隐色结晶紫，LCV）之间的差异。合理设计并合成了一个潜在的半抗原。通过免疫新西兰白兔和 BALB/C 小鼠来制备多克隆抗体。我们测试了半抗原与 TDs 之间的交叉反应率。交叉反应率与表面静电势的差异相关。兔和鼠抗体的相关系数（²）分别为 0.901 和 0.813。根据这个线性模型，原子电荷的显著差异似乎使得不可能找到一个半抗原，它可以产生与氧化和还原 TDs 都具有良好交叉反应性的抗体。
