Utiger A, Rosskopf M, Guscetti F, Ackermann M
Institute for Virology, Vet.-med. Faculty, University of Zürich.
Adv Exp Med Biol. 1993;342:197-202. doi: 10.1007/978-1-4615-2996-5_31.
We describe a new monoclonal antibody No. 204 (mcAb 204) which recognized a family of four polypeptides, consisting of a 27kD, a 24/23kD double band and a 19kD protein present within PEDV infected cell lysates. These proteins were identified by immunoprecipitation as well as by staining of immunoblots. In infected Vero cell cultures, the synthesis of the 27kD protein was initiated between 6 and 8 hours post inoculation. The 24/23kD double band and the 19kD protein were only detectable later. At least the 27 and the 24/23kD proteins were apparently glycosylated and present in purified virions. Pulse-chase as well as solubilization experiments indicated that the faster migrating bands represented processed products of the 27kD glycoprotein. The nature of the processing is not known at present. We suggest that the 27kD protein family may represent the integral membrane protein M of PEDV. Since this protein is highly abundant in virions as well as in infected cells, and since mcAb 204 is able to react with its antigen under various conditions, this monoclonal antibody may be useful to further studies of the M-protein of PEDV. In addition, it may provide a useful tool for routine diagnosis.
我们描述了一种新的单克隆抗体204(mcAb 204),它识别一组四种多肽,由27kD、24/23kD双条带和19kD蛋白质组成,存在于感染猪流行性腹泻病毒(PEDV)的细胞裂解物中。这些蛋白质通过免疫沉淀以及免疫印迹染色进行鉴定。在感染的Vero细胞培养物中,27kD蛋白质的合成在接种后6至8小时开始。24/23kD双条带和19kD蛋白质稍后才检测到。至少27kD和24/23kD蛋白质明显被糖基化并存在于纯化的病毒粒子中。脉冲追踪以及溶解实验表明,迁移较快的条带代表27kD糖蛋白的加工产物。目前尚不清楚加工的性质。我们认为27kD蛋白质家族可能代表PEDV的整合膜蛋白M。由于这种蛋白质在病毒粒子以及感染细胞中高度丰富,并且由于mcAb 204能够在各种条件下与其抗原反应,这种单克隆抗体可能有助于进一步研究PEDV的M蛋白。此外,它可能为常规诊断提供有用的工具。