Sittner Assa, Mechaly Adva, Vitner Einat, Aftalion Moshe, Levy Yinon, Levy Haim, Mamroud Emanuelle, Fisher Morly
Department of Infectious Diseases, Israel Institute for Biological Research, P.O. Box 19, Ness Ziona 74100, Israel.
Department of Biochemistry and Molecular Genetics, Israel Institute for Biological Research, P.O. Box 19, Ness Ziona 74100, Israel.
J Biol Methods. 2018 Nov 7;5(4):e100. doi: 10.14440/jbm.2018.257. eCollection 2018.
For about four decades, hybridoma technologies have been the "work horse" of monoclonal antibody production. These techniques proved to be robust and reliable, albeit laborious. Over the years, several major improvements have been introduced into the field, but yet, antibody production still requires many hours of labor and considerable resources. In this work, we present a leap forward in the advancement of hybridoma-based monoclonal antibody production, which saves labor and time and increases yield, by combining hybridoma technology, fluorescent particles and fluorescence-activated cell sorting (FACS). By taking advantage of the hybridomas' cell-surface associated antibodies, we can differentiate between antigen-specific and non-specific cells, based on their ability to bind the particles. The speed and efficiency of antibody discovery, and subsequent cell cloning, are of high importance in the field of infectious diseases. Therefore, as a model system, we chose the protein LcrV, a major virulence factor of the plague pathogen pestis, an important re-emerging pathogen and a possible bioterror agent.
在大约四十年的时间里，杂交瘤技术一直是单克隆抗体制备的“主力军”。这些技术虽被证明稳健可靠，但颇为繁琐。多年来，该领域引入了几项重大改进，但抗体生产仍需大量人工和可观的资源。在这项工作中，我们通过结合杂交瘤技术、荧光颗粒和荧光激活细胞分选（FACS），在基于杂交瘤的单克隆抗体制备方面取得了飞跃，节省了人工和时间并提高了产量。通过利用杂交瘤细胞表面相关抗体，我们可以根据细胞结合颗粒的能力区分抗原特异性细胞和非特异性细胞。抗体发现以及后续细胞克隆的速度和效率在传染病领域至关重要。因此，作为一个模型系统，我们选择了蛋白质LcrV，它是鼠疫病原体耶尔森氏菌的一种主要毒力因子，耶尔森氏菌是一种重要的再次出现的病原体，也是一种可能的生物恐怖制剂。
