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酵母杀伤毒素单克隆抗体的制备与鉴定

Production and characterization of yeast killer toxin monoclonal antibodies.

作者信息

Polonelli L, Morace G

出版信息

J Clin Microbiol. 1987 Feb;25(2):460-2. doi: 10.1128/jcm.25.2.460-462.1987.

Abstract

Monoclonal antibodies were obtained after fusion of mouse myeloma cells with spleen cells isolated from mice primed with a crude extract of yeast killer toxin produced by a strain of Hansenula anomala. Hybridomas were selected by specific immunoassay reaction of their fluid with crude yeast killer toxin extract. Among the monoclonal antibodies, which were characterized by the Western blot technique, one (designated KT4) proved to have precipitating properties, thus permitting the neutralization of the killer activity of the toxin. Experiments in double immunodiffusion showed that monoclonal antibody KT4 produced homologous precipitin bands by reacting with either the crude toxin used as immunogen or a toxic extract of Hansenula mrakii. It is suggested that these monoclonal antibodies will be useful for the purification, characterization, and understanding of the bioactions of yeast killer toxins.

摘要

将小鼠骨髓瘤细胞与用异常汉逊酵母菌株产生的酵母杀伤毒素粗提物免疫的小鼠脾脏细胞融合后,获得了单克隆抗体。通过杂交瘤细胞培养液与酵母杀伤毒素粗提物的特异性免疫测定反应来筛选杂交瘤。利用蛋白质印迹技术对单克隆抗体进行表征,其中一种(命名为KT4)具有沉淀特性,因此能够中和毒素的杀伤活性。双向免疫扩散实验表明,单克隆抗体KT4与用作免疫原的粗毒素或粟酒裂殖酵母的毒性提取物反应时,会产生同源沉淀带。这些单克隆抗体有望用于酵母杀伤毒素的纯化、表征及对其生物作用的理解。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0f0c/265928/2a5dd287feb0/jcm00086-0291-a.jpg

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