Nagakura N, Shimizu T, Masuzawa T, Yanagihara Y
Department of Microbiology, School of Pharmaceutical Sciences, University of Shizuoka, Japan.
Int Arch Allergy Appl Immunol. 1990;93(2-3):126-32. doi: 10.1159/000235291.
Three cell lines producing monoclonal antibodies (MAbs), Cep1-2, 2-2 and 6, against cephalexin were established and the immunoglobulin class of the MAbs was IgM. The cross-reactions of the MAbs with penams and cephems were examined by enzyme-linked immunosorbent assay (ELISA). The cross-reactivities of Cep1-2 and 2-2 were scarcely influenced by the structures of acyl side chains of cephems and penams. The cross-reactivities of Cep1-2 were affected by the presence or absence of dihydrothiazolidine ring of cephem nucleus in hapten-protein conjugates which were prepared by alkaline method, MBS method and activated ester method but the cross-reactivities of Cep2-2 were not. The findings suggest that Cep1-2 recognize the degradate product(s) of cephem nucleus and Cep2-2 recognize a new antigenic determinant (NAD), which is formed by the conjugation of beta-lactam and carrier protein. On the other hand, the cross-reactivities of Cep6 were influenced by the structure of amino acyl side chain. It seems that Cep6 recognize specifically the acyl side chain at the C-7 of cephem. In ELISA inhibition test, three MAbs showed different inhibition pattern. The reaction of Cep1-2 with cephalexin-HSA was inhibited by cephalexin lysate. Cep2-2 and Cep6 were weakly inhibited by the binding to cephalexin-HSA by cephalexin lysate. Furthermore, the reactions of all MAbs were remarkably inhibited by penicillamine. The above results indicate that the MAbs can recognize at least three epitopes of the degradate product(s) of cephem nucleus, NAD and acyl side chain in cephalexin-protein conjugate.
建立了三种产生抗头孢氨苄单克隆抗体（MAb）的细胞系，即Cep1-2、2-2和6，这些单克隆抗体的免疫球蛋白类别为IgM。通过酶联免疫吸附测定（ELISA）检测了单克隆抗体与青霉烷和头孢菌素的交叉反应。Cep1-2和2-2的交叉反应几乎不受头孢菌素和青霉烷酰基侧链结构的影响。通过碱性方法、MBS方法和活化酯方法制备的半抗原-蛋白质缀合物中，头孢菌素核二氢噻唑烷环的有无会影响Cep1-2的交叉反应，但不会影响Cep2-2的交叉反应。这些发现表明，Cep1-2识别头孢菌素核的降解产物，而Cep2-2识别一种新的抗原决定簇（NAD），它是由β-内酰胺与载体蛋白结合形成的。另一方面，Cep6的交叉反应受氨基酰基侧链结构的影响。似乎Cep6特异性识别头孢菌素C-7位的酰基侧链。在ELISA抑制试验中，三种单克隆抗体表现出不同的抑制模式。Cep1-2与头孢氨苄-HSA的反应被头孢氨苄裂解物抑制。头孢氨苄裂解物与头孢氨苄-HSA的结合对Cep2-2和Cep6有较弱的抑制作用。此外，所有单克隆抗体的反应都被青霉胺显著抑制。上述结果表明，单克隆抗体可以识别头孢氨苄-蛋白质缀合物中头孢菌素核降解产物、NAD和酰基侧链的至少三个表位。
