Generating a monoclonal antibody to date is a time intense process that requires immunization of laboratory animals. The transfer of the humoral immune response into in vitro settings enables a shortening of this process and circumvents the necessity of in vivo immunization. However, to orchestrate the complex interplay of dendritic cells, T and B lymphocytes in vitro is very challenging. We therefore aimed for a simplified approach focusing on the protagonist of antibody production: the B lymphocyte. We activated purified murine B lymphocytes alone in vitro by using combinations of antigen and stimuli. We were able to induce a specific antibody response within ten days of culture against a viral coat protein as model antigen. Antibodies were of both IgM and IgG subclass. The stimulated B lymphocytes were transformed into permanently antibody-producing hybridomas by cell fusion technology. We furthermore used this method to induce a specific antibody response against L. pneumophila in vitro. We thus established a useful and effective in vitro protocol to generate monoclonal antibodies. By overcoming the necessity of in vivo immunization this protocol may be the first step towards a universal strategy to generate antibodies from various species.

迄今为止，产生单克隆抗体是一个时间紧迫的过程，需要对实验动物进行免疫接种。将体液免疫反应转移到体外环境中可以缩短这一过程并避免体内免疫的必要性。然而，要在体外协调树突状细胞、T 和 B 淋巴细胞的复杂相互作用是非常具有挑战性的。因此，我们旨在采用一种简化方法，重点关注抗体生产的主角：B 淋巴细胞。我们在体外单独激活纯化的鼠 B 淋巴细胞通过使用抗原和刺激物的组合。我们能够在培养十天内针对作为模型抗原的病毒外壳蛋白诱导特异性抗体反应。抗体属于 IgM 和 IgG 亚类。通过细胞融合技术将受刺激的 B 淋巴细胞转化为永久产生抗体的杂交瘤。我们进一步使用这种方法在体外诱导针对嗜肺军团菌 的特异性抗体反应。因此，我们建立了一个有用且有效的体外方案来生成单克隆抗体。通过克服体内免疫的必要性，该协议可能是迈向从不同物种产生抗体的通用策略的第一步。