Utz U, Britt W, Vugler L, Mach M
Institut für Klinische und Molekulare Virologie, Universität Erlangen-Nürnberg, Federal Republic of Germany.
J Virol. 1989 May;63(5):1995-2001. doi: 10.1128/JVI.63.5.1995-2001.1989.
Human cytomegalovirus contains an envelope glycoprotein of 58 kilodaltons (gp58). The protein, which is derived from a glycosylated precursor molecule of 160 kilodaltons via proteolytic cleavage, is capable of inducing neutralizing antibodies. We have mapped the epitopes recognized by the neutralizing monoclonal antibody 7-17 and a second antibody (27-287) which is not neutralizing. Overlapping fragments of the carboxy-terminal part of the open reading frame coding for gp58 were expressed in Escherichia coli as beta-galactosidase fusion proteins. The reactivities of antibodies 7-17 and 27-287 were determined by Western blot (immunoblot) analysis. Both antibodies recognized sequences between amino acids 608 and 625 of the primary gp58 translation product. The antibodies almost completely inhibited one another in a competitive binding assay with intact virus as antigen. Moreover, antibody 27-287 was able to inhibit the complement-independent neutralizing activity of antibody 7-17.
人巨细胞病毒含有一种58千道尔顿的包膜糖蛋白（gp58）。该蛋白由一个160千道尔顿的糖基化前体分子经蛋白水解切割产生，能够诱导中和抗体。我们已经绘制了中和单克隆抗体7 - 17和第二种非中和抗体（27 - 287）所识别的表位。编码gp58的开放阅读框羧基末端部分的重叠片段在大肠杆菌中作为β - 半乳糖苷酶融合蛋白表达。通过蛋白质印迹（免疫印迹）分析确定抗体7 - 17和27 - 287的反应性。两种抗体都识别初级gp58翻译产物氨基酸608至625之间的序列。在以完整病毒为抗原的竞争性结合试验中，两种抗体几乎完全相互抑制。此外，抗体27 - 287能够抑制抗体7 - 17的非补体依赖性中和活性。
