Comprehensive Method for Producing High-affinity Mouse Monoclonal Antibodies of Various Isotypes Against (4-hydroxy-3-nitrophenyl)acetyl (NP) Hapten

Overview

Journal Heliyon

Specialty Social Sciences

Date 2024 Dec 19

PMID 39698082

Authors

Rin Yoshizato

Mariko Miura

Kiyomi Shitaoka

Yuri Matsuoka

Akifumi Higashiura

Akima Yamamoto

Yun Guo

Hitoshi Azuma

Yohei Kawano

Shouichi Ohga

Tomoharu Yasuda

Affiliations

Soon will be listed here.

Abstract

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.

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