Expression Analysis of Heavy-Chain-Only Antibodies in Cloudy Catshark and Japanese Bullhead Shark
Heavy chain-only antibodies in sharks are called immunoglobulin new antigen receptors (IgNAR), consisting of one variable region (VNAR) and five constant regions (C1-C5). The variable region of IgNAR can be expressed as a monomer composed of a single domain, which has antigen specificity and is thus...
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2025-01-01
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| Series: | Marine Drugs |
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| author | Reo Uemura Susumu Tanimura Nao Yamaguchi Ryuichi Kuroiwa Gabriel Takashi Andrés Tsutsumi Toshiaki Fujikawa Kiyoshi Soyano Kohsuke Takeda Yoshimasa Tanaka |
| author_facet | Reo Uemura Susumu Tanimura Nao Yamaguchi Ryuichi Kuroiwa Gabriel Takashi Andrés Tsutsumi Toshiaki Fujikawa Kiyoshi Soyano Kohsuke Takeda Yoshimasa Tanaka |
| author_sort | Reo Uemura |
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| description | Heavy chain-only antibodies in sharks are called immunoglobulin new antigen receptors (IgNAR), consisting of one variable region (VNAR) and five constant regions (C1-C5). The variable region of IgNAR can be expressed as a monomer composed of a single domain, which has antigen specificity and is thus gaining attention as a next-generation antibody drug modality. In this study, we analyzed IgNAR of the cloudy catshark and Japanese bullhead shark, small demersal sharks available in the coastal waters of Japan. By analyzing the IgNAR gene sequence and comparing it with the constant regions of five other known shark species, high homology was observed in the C4 region. Consequently, we expressed the recombinant protein of the C4 domain from the cloudy catshark in <i>E. coli</i>, immunized rats, and produced antibodies. The obtained antiserum and mAbs recognized the C4 recombinant protein of the cloudy catshark, but reacted minimally with the plasma of non-immunized cloudy catsharks and instead reacted with the plasma of Japanese bullhead sharks. The results of this study imply that the protein expression levels of IgNAR in cloudy catsharks may be relatively lower compared to those in Japanese bullhead sharks, however, this interpretation remains to be determined through further studies. |
| format | Article |
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| institution | Kabale University |
| issn | 1660-3397 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | MDPI AG |
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| series | Marine Drugs |
| spelling | doaj-art-f7163ffaaaf948f2bb4ce99b47e9a9132025-01-24T13:39:32ZengMDPI AGMarine Drugs1660-33972025-01-012312810.3390/md23010028Expression Analysis of Heavy-Chain-Only Antibodies in Cloudy Catshark and Japanese Bullhead SharkReo Uemura0Susumu Tanimura1Nao Yamaguchi2Ryuichi Kuroiwa3Gabriel Takashi Andrés Tsutsumi4Toshiaki Fujikawa5Kiyoshi Soyano6Kohsuke Takeda7Yoshimasa Tanaka8Department of Cell Regulation, Graduate School of Biomedical Sciences, Nagasaki University, Bunkyo-machi 1-14, Nagasaki 852-8521, JapanDepartment of Cell Regulation, Graduate School of Biomedical Sciences, Nagasaki University, Bunkyo-machi 1-14, Nagasaki 852-8521, JapanDepartment of Cell Regulation, Graduate School of Biomedical Sciences, Nagasaki University, Bunkyo-machi 1-14, Nagasaki 852-8521, JapanDepartment of Cell Regulation, Graduate School of Biomedical Sciences, Nagasaki University, Bunkyo-machi 1-14, Nagasaki 852-8521, JapanDepartment of Cell Regulation, Graduate School of Biomedical Sciences, Nagasaki University, Bunkyo-machi 1-14, Nagasaki 852-8521, JapanInstitute for East China Sea Research, Organization for Marine Science and Technology, Nagasaki University, 1551-7, Taira-machi, Nagasaki 851-2213, JapanInstitute for East China Sea Research, Organization for Marine Science and Technology, Nagasaki University, 1551-7, Taira-machi, Nagasaki 851-2213, JapanDepartment of Cell Regulation, Graduate School of Biomedical Sciences, Nagasaki University, Bunkyo-machi 1-14, Nagasaki 852-8521, JapanCenter for Medical Innovation, Nagasaki University, Sakamoto 1-7-1, Nagasaki 852-8588, JapanHeavy chain-only antibodies in sharks are called immunoglobulin new antigen receptors (IgNAR), consisting of one variable region (VNAR) and five constant regions (C1-C5). The variable region of IgNAR can be expressed as a monomer composed of a single domain, which has antigen specificity and is thus gaining attention as a next-generation antibody drug modality. In this study, we analyzed IgNAR of the cloudy catshark and Japanese bullhead shark, small demersal sharks available in the coastal waters of Japan. By analyzing the IgNAR gene sequence and comparing it with the constant regions of five other known shark species, high homology was observed in the C4 region. Consequently, we expressed the recombinant protein of the C4 domain from the cloudy catshark in <i>E. coli</i>, immunized rats, and produced antibodies. The obtained antiserum and mAbs recognized the C4 recombinant protein of the cloudy catshark, but reacted minimally with the plasma of non-immunized cloudy catsharks and instead reacted with the plasma of Japanese bullhead sharks. The results of this study imply that the protein expression levels of IgNAR in cloudy catsharks may be relatively lower compared to those in Japanese bullhead sharks, however, this interpretation remains to be determined through further studies.https://www.mdpi.com/1660-3397/23/1/28cloudy catsharkheavy-chain-only antibodyimmunoglobulin new antigen receptorJapanese bullhead sharkVNAR-domain antibody |
| spellingShingle | Reo Uemura Susumu Tanimura Nao Yamaguchi Ryuichi Kuroiwa Gabriel Takashi Andrés Tsutsumi Toshiaki Fujikawa Kiyoshi Soyano Kohsuke Takeda Yoshimasa Tanaka Expression Analysis of Heavy-Chain-Only Antibodies in Cloudy Catshark and Japanese Bullhead Shark Marine Drugs cloudy catshark heavy-chain-only antibody immunoglobulin new antigen receptor Japanese bullhead shark VNAR-domain antibody |
| title | Expression Analysis of Heavy-Chain-Only Antibodies in Cloudy Catshark and Japanese Bullhead Shark |
| title_full | Expression Analysis of Heavy-Chain-Only Antibodies in Cloudy Catshark and Japanese Bullhead Shark |
| title_fullStr | Expression Analysis of Heavy-Chain-Only Antibodies in Cloudy Catshark and Japanese Bullhead Shark |
| title_full_unstemmed | Expression Analysis of Heavy-Chain-Only Antibodies in Cloudy Catshark and Japanese Bullhead Shark |
| title_short | Expression Analysis of Heavy-Chain-Only Antibodies in Cloudy Catshark and Japanese Bullhead Shark |
| title_sort | expression analysis of heavy chain only antibodies in cloudy catshark and japanese bullhead shark |
| topic | cloudy catshark heavy-chain-only antibody immunoglobulin new antigen receptor Japanese bullhead shark VNAR-domain antibody |
| url | https://www.mdpi.com/1660-3397/23/1/28 |
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