Rohde M, Schenk J A, Heymann S, Behrsing O, Scharte G, Kempter G, Woller J, Höhne W E, Warsinke A, Micheel B
Potsdam University, Institute of Biochemistry and Molecular Physiology, Berlin.
Appl Biochem Biotechnol. 1998 Oct;75(1):129-37. doi: 10.1007/BF02787713.
A panel of monoclonal antibodies was generated against the urea-based hapten N-(2-N-chloroacetylaminobenzyl)-N'-4-chlorophenylurea as a tool for building up sensitive immune assays to detect urea derivatives and to screen them for catalytic antibodies (Abs). Eleven hybridomas were obtained that produced Abs reactive to the hapten. All Abs were of IgG class. Cross reactivities of the Abs to different haptens were examined, especially to a possible transition-state analog. Only four of the hybridomas (R2-DA10/F7, R2-GE7/H2, R2-HC2/A5, R2-HD6/F7) produced Abs crossreactive with the transition-state analog. From the 11 hybridomas, hybridoma B76-BF5 was chosen for further characterization. Compared to the other Abs, B76-BF5 showed the strongest binding and had a rather restricted specificity. These Abs could be used to build up a sensitive enzyme immunoassay for the detection of the hapten. All Abs were screened for crossreactivity with the pesticides monuron and diuron. No reactivity could be detected. In addition, the nucleotide sequences of the variable light and heavy chain genes of the similarly reactive Abs B76-BF5, B76-BB3, R2-DA10/F7, and R2-GA6/G3 were determined to clarify whether structure and binding specificity of these Abs showed any correlation.
制备了一组针对基于尿素的半抗原N-(2-N-氯乙酰氨基苄基)-N'-4-氯苯基脲的单克隆抗体，作为建立灵敏免疫测定法以检测尿素衍生物并筛选催化抗体（Abs）的工具。获得了11个产生对半抗原具有反应性的抗体的杂交瘤。所有抗体均为IgG类。检测了这些抗体对不同半抗原的交叉反应性，特别是对可能的过渡态类似物的交叉反应性。只有四个杂交瘤（R2-DA10/F7、R2-GE7/H2、R2-HC2/A5、R2-HD6/F7）产生了与过渡态类似物交叉反应的抗体。从这11个杂交瘤中，选择杂交瘤B76-BF5进行进一步表征。与其他抗体相比，B76-BF5表现出最强的结合力且特异性相当有限。这些抗体可用于建立检测半抗原的灵敏酶免疫测定法。检测了所有抗体与农药灭草隆和敌草隆的交叉反应性。未检测到反应性。此外，测定了具有相似反应性的抗体B76-BF5、B76-BB3、R2-DA10/F7和R2-GA6/G3的可变轻链和重链基因的核苷酸序列，以阐明这些抗体的结构和结合特异性是否存在任何相关性。
