Academic Journal
Comprehensive method for producing high-affinity mouse monoclonal antibodies of various isotypes against (4-hydroxy-3-nitrophenyl)acetyl (NP) hapten
| Title: | Comprehensive method for producing high-affinity mouse monoclonal antibodies of various isotypes against (4-hydroxy-3-nitrophenyl)acetyl (NP) hapten |
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| Authors: | Rin Yoshizato, Mariko Miura, Kiyomi Shitaoka, Yuri Matsuoka, Akifumi Higashiura, Akima Yamamoto, Yun Guo, Hitoshi Azuma, Yohei Kawano, Shouichi Ohga, Tomoharu Yasuda |
| Source: | Heliyon, Vol 10, Iss 23, Pp e40837- (2024) |
| Publisher Information: | Elsevier, 2024. |
| Publication Year: | 2024 |
| Collection: | LCC:Science (General) LCC:Social sciences (General) |
| Subject Terms: | Monoclonal antibody, Memory B cells, B cell receptor, Single cell, Affinity maturation, IgM, Science (General), Q1-390, Social sciences (General), H1-99 |
| More Details: | 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. |
| Document Type: | article |
| File Description: | electronic resource |
| Language: | English |
| ISSN: | 2405-8440 |
| Relation: | http://www.sciencedirect.com/science/article/pii/S2405844024168685; https://doaj.org/toc/2405-8440 |
| DOI: | 10.1016/j.heliyon.2024.e40837 |
| Access URL: | https://doaj.org/article/d9668eb981594ab18c46ba8ed36326df |
| Accession Number: | edsdoj.9668eb981594ab18c46ba8ed36326df |
| Database: | Directory of Open Access Journals |
| ISSN: | 24058440 |
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| DOI: | 10.1016/j.heliyon.2024.e40837 |
| Published in: | Heliyon |
| Language: | English |