Beier R C, Ripley L H, Young C R, Kaiser C M
Southern Plains Agricultural Research Center, Agricultural Research Service, U.S. Department of Agriculture, 2881 F & B Road, College Station, Texas 77845-4988, USA.
J Agric Food Chem. 2001 Oct;49(10):4542-52. doi: 10.1021/jf010208j.
A cELISA was developed for the coccidiostat nicarbazin. On the basis of previous computer-assisted molecular modeling studies, p-nitrosuccinanilic acid (PNA-S) was selected as a hapten to produce antibodies to 4,4'-dinitrocarbanilide (DNC), the active component of the coccidiostat nicarbazin. Synthesis is described for the hapten [p-nitro-cis-1,2-cyclohexanedicarboxanilic acid (PNA-C)] used in a BSA conjugate as a plate coating antigen. Monoclonal antibodies (Mabs) were isolated that compete with nicarbazin, having IgM(kappa) isotype. Because of the lack of water solubility of nicarbazin, N,N-dimethylformamide (DMF) (3%, v/v) and acetonitrile (ACN) (10%, v/v) were added to the assay buffer to achieve solubility of nicarbazin and related compounds. The Nic 6 Mabs had an IC(35) value for nicarbazin of 0.92 nmol/mL, with a limit of detection of 0.33 nmol/mL. Nic 6 exhibited high cross-reactivity for PNA-S and PNA-C, and 3-nitrophenol, 4-nitrophenol, and 1-(4-chlorophenyl)-3-(4-nitrophenyl) urea. However, Nic 6 had little or no cross-reactivity with 15 other related compounds.
开发了一种用于抗球虫药尼卡巴嗪的竞争酶联免疫吸附测定（cELISA）。基于先前的计算机辅助分子建模研究，选择对硝基琥珀酰苯胺（PNA-S）作为半抗原，以产生针对抗球虫药尼卡巴嗪的活性成分4,4'-二硝基碳酰苯胺（DNC）的抗体。描述了用于牛血清白蛋白（BSA）偶联物作为板包被抗原的半抗原[对硝基-顺式-1,2-环己烷二甲酰苯胺（PNA-C）]的合成。分离出与尼卡巴嗪竞争的单克隆抗体（Mab），其具有IgM（κ）同种型。由于尼卡巴嗪缺乏水溶性，在测定缓冲液中加入N，N-二甲基甲酰胺（DMF）（3%，v/v）和乙腈（ACN）（10%，v/v）以实现尼卡巴嗪及相关化合物的溶解。Nic 6单克隆抗体对尼卡巴嗪的IC(35)值为0.92 nmol/mL，检测限为0.33 nmol/mL。Nic 6对PNA-S、PNA-C、3-硝基苯酚、4-硝基苯酚和1-(4-氯苯基)-3-(4-硝基苯基)脲表现出高交叉反应性。然而，Nic 6与其他15种相关化合物几乎没有交叉反应性。
