Yoshizato Rin, Miura Mariko, Shitaoka Kiyomi, Matsuoka Yuri, Higashiura Akifumi, Yamamoto Akima, Guo Yun, Azuma Hitoshi, Kawano Yohei, Ohga Shouichi, Yasuda Tomoharu
Department of Immunology, Graduate School of Biomedical and Health Sciences, Hiroshima University, Hiroshima, Japan.
Department of Pediatrics, Graduate School of Medical Sciences, Kyushu University, Fukuoka, Japan.
Heliyon. 2024 Nov 29;10(23):e40837. doi: 10.1016/j.heliyon.2024.e40837. eCollection 2024 Dec 15.
Monoclonal antibody (mAb) technology has significantly contributed to basic research and clinical settings for various purposes, including protective and therapeutic drugs. However, a rapid and convenient method to generate high-affinity antigen-specific mAbs has not yet been reported. Here, we developed a rapid, easy, and low-cost protocol for antigen-specific mAb production from single memory B cells. Using this method, high-affinity IgG1 mAbs specific to the hapten 4-hydroxy-3-nitrophenylacetyl (NP) were established from NP-CGG immunized C57BL/6 mice within 6 days. Our mAb production system allows flexible switching of IgG1 to any other isotype with the same paratope, enabling the absolute quantification of antigen-specific serum antibody titers and affinity maturation. Additionally, we established a protocol for the production of IgM and IgA, retaining their functional pentamer and dimer structures. This method is also effective against human antigens and pathogens, making it a powerful tool for mAb development in both research and clinical settings.
单克隆抗体（mAb）技术在包括保护性和治疗性药物在内的各种基础研究和临床应用中都做出了重大贡献。然而，尚未有报道一种快速便捷的方法来产生高亲和力的抗原特异性单克隆抗体。在此，我们开发了一种从单个记忆B细胞产生抗原特异性单克隆抗体的快速、简便且低成本的方案。使用该方法，在6天内从经NP-CGG免疫的C57BL/6小鼠中建立了对半抗原4-羟基-3-硝基苯乙酰（NP）具有特异性的高亲和力IgG1单克隆抗体。我们的单克隆抗体制备系统允许将IgG1灵活切换为具有相同抗原结合位点的任何其他同种型，能够绝对定量抗原特异性血清抗体滴度并实现亲和力成熟。此外，我们建立了保留其功能性五聚体和二聚体结构的IgM和IgA的制备方案。该方法对人类抗原和病原体也有效，使其成为研究和临床环境中单克隆抗体开发的有力工具。
