New screening method for fish monoclonal antibody gene and establishment of antibody expression system
| Project/Area Number |
15H02461
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| Research Category |
Grant-in-Aid for Scientific Research (A)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Aquatic life science
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| Research Institution | The University of Tokyo |
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Principal Investigator |
Asakawa Shuichi 東京大学, 大学院農学生命科学研究科(農学部), 教授 (30231872)
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| Co-Investigator(Kenkyū-buntansha) |
大政 健史 大阪大学, 工学(系)研究科(研究院), 准教授 (00252586)
近藤 秀裕 東京海洋大学, 学術研究院, 准教授 (20314635)
田角 聡志 東京大学, 大学院農学生命科学研究科(農学部), 特任助教 (90359646)
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| Project Period (FY) |
2015-04-01 – 2018-03-31
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| Project Status |
Completed (Fiscal Year 2017)
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| Budget Amount *help |
¥42,250,000 (Direct Cost: ¥32,500,000、Indirect Cost: ¥9,750,000)
Fiscal Year 2017: ¥13,000,000 (Direct Cost: ¥10,000,000、Indirect Cost: ¥3,000,000)
Fiscal Year 2016: ¥13,000,000 (Direct Cost: ¥10,000,000、Indirect Cost: ¥3,000,000)
Fiscal Year 2015: ¥16,250,000 (Direct Cost: ¥12,500,000、Indirect Cost: ¥3,750,000)
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| Keywords | IgNAR / 一本鎖抗体 / サメ抗体 / 免疫クロブリン / トランスクリプトーム / CHO細胞 / VLR / トラフグ / イヌザメ / ドチザメ / 抗体医薬 / 人工抗体 / 水産学 / 生物工学 / バイオインフォーマティクス / ゲノム / モノクローナル抗体 / 抗体工学 / 水圏生物工学 / サメ / ヤツメウナギ |
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| Outline of Final Research Achievements |
The cDNA sequence of IgNAR of bamboo shark (Chiloscyllium punctatum) was obtained and the sequence of the constant region was extracted. We also established a system to amplify the variable region of IgNAR and measure the expression frequency of individual genes.
Antigens (HEL, BSA, MMP-09) were administered several times to the bamboo shark at intervals of one month, and blood was collected one month after administration. Antibody titer was measured by ELISA for BSA and multiple genes with increased expression frequency were identified in conjunction with rise in antibody titer. Expression constructs combining variable and constant regions of shark antibodies were prepared using CHO cells and succeeded in secretory production. From these results, identification of the antigen-specific antibody gene by transcriptome and establishment of its expression system were almost completed.
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Report
(4 results)
Research Products
(19 results)
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[Journal Article] Ultrahigh-Density Linkage Map Construction Using Low-Coverage Whole-Genome Sequencing of a Doubled Haploid Population: Case Study of Torafugu (Takifugu rubripes).2018
Author(s)
Zhang X, Mizukoshi M, Zhang H, Tan E, Igarashi Y, Suzuki Y, Mitsuyama S, Kinoshita S, Saito K, Watabe S, Asakawa S
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Journal Title
Genes
Volume: 9
Issue: 3
Pages: 120-120
DOI
Related Report
Open Access
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[Presentation] Determination of immunoglobulin novel antigen receptor (IgNAR) in vivo affinity maturation in brownbanded bamboo shark (Chiloscyllium punctatum).2017
Author(s)
2.De Silva, D.P.N., Hosoya, S., Mizumo, N., Kinoshita, S., Totsuka, M. and Asakawa, S.
Organizer
The JSFS 85th Anniversary-Commemorative International Symposium
Related Report
Int'l Joint Research
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[Presentation] Determination of variable region of novel antigen receptor (vNAR) from brownbanded bamboo shark (Chiloscyllium punctatum).2016
Author(s)
2.De Silva, D.P.N., Tan, E., Mizumo, N., Kinoshita, S., Mitsuyama, S. And Asakawa, S.
Organizer
7th World Fisheries Congress
Place of Presentation
Busan, Korea
Year and Date
2016-05-23
Related Report
Int'l Joint Research
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[Presentation] reliminary investigation of brownbanded bamboo shark (Chiloscyllium punctatum) immunoglobulin light chain isotypes.2016
Author(s)
De Silva, D.P.N., Tan, E., Mizumo, N., Kinoshita, S., Mitsuyama, S. And Asakawa, S.
Organizer
7th World Fisheries Congress
Place of Presentation
Busan, Korea
Year and Date
2016-05-23
Related Report
Int'l Joint Research
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[Presentation] Preliminary identification of immunoglobulin isotypes in brownbanded bamboo shark (Chiloscyllium punctatum) by heavy chain constant domain analysis2015
Author(s)
2.De Silva, D.P.N., Tan, E., Mizumo, N., Kinoshita, S., Mitsuyama, S. And Asakawa, S.
Organizer
第38回日本分子生物学会・第88回日本生化学会合同大会（BMB2015）
Place of Presentation
神戸（神戸ポートアイランド）
Year and Date
2015-12-01
Related Report
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[Presentation] Characterization of the immunoglobulin novel antigen receptor gene of brownbanded bamboo shark (Chiloscyllium punctatum).2015
Author(s)
1.De Silva, D.P.N., Tan, E., Mizumo, N., Kinoshita, S., Mitsuyama, S. And Asakawa, S.
Organizer
平成27年度日本水産学会秋季大会
Place of Presentation
仙台（東北大学川内北キャンパス）
Year and Date
2015-09-22
Related Report
