The purpose of this work was to obtain genus-specific monoclonal antibodies against the Legionella spp. recombinant PAL protein, which will subsequently allow to use them as a basis for the development of new express tests for pathogenic legionella detection. A short three-week immunization protocol for Wistar rats was used to generate rat–mouse heterohybridomas producing antibodies against PAL. Mouse myeloma cell line Sp2/0-Ag14 served as the fusion partner. Hybridization was performed using two methods: PEG-mediated fusion and electrofusion. Subsequent screening was performed by indirect solid-phase ELISA against the target protein rPAL. Specificity analysis was performed by dot-blot using a panel of lysates obtained from 39 pure cultures of different strains, which included closely related and heterologous microorganisms among others. No difference in the efficiency of stable hybridoma clones production by the two indicated cell-fusion methods was detected. Twelve clones producing specific rat monoclonal antibodies were obtained based on the screening results. The obtained rat monoclonal antibodies are highly specific towards the PAL protein of L. pneumophila of different serological groups and other pathogenic legionella and are good candidates to be used as the components of diagnostic test systems for the detection of pathogenic representatives of the Legionella genus.

这项工作的目的是获得针对军团菌的属特异性单克隆抗体spp. 重组 PAL 蛋白，随后将允许将它们用作开发用于检测致病性军团菌的新快速测试的基础。对 Wistar 大鼠进行了为期三周的短免疫方案，用于产生产生抗 PAL 抗体的大鼠 - 小鼠异种杂交瘤。小鼠骨髓瘤细胞系 Sp2/0-Ag14 作为融合伙伴。使用两种方法进行杂交：PEG介导的融合和电融合。随后通过针对靶蛋白rPAL的间接固相ELISA进行筛选。使用从 39 种不同菌株的纯培养物中获得的一组裂解物通过斑点印迹进行特异性分析，其中包括密切相关的和异源微生物等。没有检测到通过两种指定的细胞融合方法产生稳定杂交瘤克隆的效率有差异。根据筛选结果，获得了12个产生特异性大鼠单克隆抗体的克隆。获得的大鼠单克隆抗体对PAL蛋白具有高度特异性L. pneumophila是不同血清学组和其他致病性军团菌的良好候选者，可用作检测军团菌属致病性代表的诊断测试系统的组件。