Institute of Biochemistry and Biology, University of Potsdam, Karl-Liebknecht-Str. 24-25, D-14476 Potsdam, Germany.
J Immunol Methods. 2021 Dec;499:113149. doi: 10.1016/j.jim.2021.113149. Epub 2021 Sep 21.
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. pneumophilain 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.
目前，产生单克隆抗体是一个时间密集型的过程，需要对实验动物进行免疫。将体液免疫反应转移到体外环境中可以缩短这个过程，并避免体内免疫的必要性。然而，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的方法，专注于抗体产生的主角：B 淋巴细胞。我们通过使用抗原和刺激物的组合，在体外单独激活纯化的小鼠 B 淋巴细胞。我们能够在 10 天的培养期内诱导针对病毒外壳蛋白的特异性抗体反应，作为模型抗原。抗体为 IgM 和 IgG 亚类。通过细胞融合技术，刺激的 B 淋巴细胞被转化为永久性产生抗体的杂交瘤。我们还使用这种方法在体外诱导针对肺炎支原体的特异性抗体反应。因此，我们建立了一种有用且有效的体外方案来产生单克隆抗体。通过克服体内免疫的必要性，该方案可能是针对各种物种产生抗体的通用策略的第一步。
