Development of a new genotype–phenotype linked antibody screening system
Antibodies are powerful tools for the therapy and diagnosis of various diseases. In addition to conventional hybridoma-based screening, recombinant antibody-based screening has become a common choice; however, its application is hampered by two factors: (1) screening starts after Ig gene cloning and...
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| Main Authors: | , , , , , , , , |
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| Format: | Article |
| Language: | English |
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eLife Sciences Publications Ltd
2024-11-01
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| Series: | eLife |
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| Online Access: | https://elifesciences.org/articles/95346 |
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| _version_ | 1846163427000582144 |
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| author | Takashi Watanabe Hikaru Hata Yoshiki Mochizuki Fumie Yokoyama Tomoko Hasegawa Naveen Kumar Tomohiro Kurosaki Osamu Ohara Hidehiro Fukuyama |
| author_facet | Takashi Watanabe Hikaru Hata Yoshiki Mochizuki Fumie Yokoyama Tomoko Hasegawa Naveen Kumar Tomohiro Kurosaki Osamu Ohara Hidehiro Fukuyama |
| author_sort | Takashi Watanabe |
| collection | DOAJ |
| description | Antibodies are powerful tools for the therapy and diagnosis of various diseases. In addition to conventional hybridoma-based screening, recombinant antibody-based screening has become a common choice; however, its application is hampered by two factors: (1) screening starts after Ig gene cloning and recombinant antibody production only, and (2) the antibody is composed of paired chains, heavy and light, commonly expressed by two independent expression vectors. Here, we introduce a method for the rapid screening of recombinant monoclonal antibodies by establishing a Golden Gate-based dual-expression vector and in-vivo expression of membrane-bound antibodies. Using this system, we demonstrate the rapid isolation of influenza cross-reactive antibodies with high affinity from immunized mice within 7 days. This system is particularly useful for isolating therapeutic or diagnostic antibodies, for example during foreseen pandemics. |
| format | Article |
| id | doaj-art-7c8635ab87ed41a1accabbc80ff78a36 |
| institution | Kabale University |
| issn | 2050-084X |
| language | English |
| publishDate | 2024-11-01 |
| publisher | eLife Sciences Publications Ltd |
| record_format | Article |
| series | eLife |
| spelling | doaj-art-7c8635ab87ed41a1accabbc80ff78a362024-11-19T07:30:23ZengeLife Sciences Publications LtdeLife2050-084X2024-11-011310.7554/eLife.95346Development of a new genotype–phenotype linked antibody screening systemTakashi Watanabe0https://orcid.org/0000-0002-6184-0375Hikaru Hata1Yoshiki Mochizuki2Fumie Yokoyama3Tomoko Hasegawa4Naveen Kumar5Tomohiro Kurosaki6Osamu Ohara7https://orcid.org/0000-0002-3328-9571Hidehiro Fukuyama8https://orcid.org/0000-0002-6457-0630Laboratory for Integrative Genomics, RIKEN Center for Integrative Medical Sciences, Yokohama, JapanLaboratory for Lymphocyte Differentiation, RIKEN Center for Integrative Medical Sciences, Yokohama, JapanLaboratory for Integrative Genomics, RIKEN Center for Integrative Medical Sciences, Yokohama, JapanLaboratory for Integrative Genomics, RIKEN Center for Integrative Medical Sciences, Yokohama, JapanLaboratory for Integrative Genomics, RIKEN Center for Integrative Medical Sciences, Yokohama, JapanLaboratory for Integrated Bioinformatics, RIKEN Center for Integrative Medical Sciences, Yokohama, JapanLaboratory for Lymphocyte Differentiation, RIKEN Center for Integrative Medical Sciences, Yokohama, Japan; Laboratory of Lymphocyte Differentiation, WPI Immunology Frontier Research Center, Osaka University, Osaka, JapanLaboratory for Integrative Genomics, RIKEN Center for Integrative Medical Sciences, Yokohama, Japan; Department of Applied Genomics, Kazusa DNA Research Institute, Chiba, JapanLaboratory for Lymphocyte Differentiation, RIKEN Center for Integrative Medical Sciences, Yokohama, Japan; Division of Immunology, Near-InfraRed Photo-Immunotherapy Research Institute, Kansai Medical University, Osaka, Japan; INSERM EST, Strasbourg Cedex, FranceAntibodies are powerful tools for the therapy and diagnosis of various diseases. In addition to conventional hybridoma-based screening, recombinant antibody-based screening has become a common choice; however, its application is hampered by two factors: (1) screening starts after Ig gene cloning and recombinant antibody production only, and (2) the antibody is composed of paired chains, heavy and light, commonly expressed by two independent expression vectors. Here, we introduce a method for the rapid screening of recombinant monoclonal antibodies by establishing a Golden Gate-based dual-expression vector and in-vivo expression of membrane-bound antibodies. Using this system, we demonstrate the rapid isolation of influenza cross-reactive antibodies with high affinity from immunized mice within 7 days. This system is particularly useful for isolating therapeutic or diagnostic antibodies, for example during foreseen pandemics.https://elifesciences.org/articles/95346membrane-bound Ig expressionngs-based Ig-seqgolden gate cloninginfluenza broadly-reactive |
| spellingShingle | Takashi Watanabe Hikaru Hata Yoshiki Mochizuki Fumie Yokoyama Tomoko Hasegawa Naveen Kumar Tomohiro Kurosaki Osamu Ohara Hidehiro Fukuyama Development of a new genotype–phenotype linked antibody screening system eLife membrane-bound Ig expression ngs-based Ig-seq golden gate cloning influenza broadly-reactive |
| title | Development of a new genotype–phenotype linked antibody screening system |
| title_full | Development of a new genotype–phenotype linked antibody screening system |
| title_fullStr | Development of a new genotype–phenotype linked antibody screening system |
| title_full_unstemmed | Development of a new genotype–phenotype linked antibody screening system |
| title_short | Development of a new genotype–phenotype linked antibody screening system |
| title_sort | development of a new genotype phenotype linked antibody screening system |
| topic | membrane-bound Ig expression ngs-based Ig-seq golden gate cloning influenza broadly-reactive |
| url | https://elifesciences.org/articles/95346 |
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