| Project/Area Number |
25460657
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Multi-year Fund |
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| Section | 一般 |
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| Research Field |
Applied pharmacology
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| Research Institution | Okayama Prefectural University |
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Principal Investigator |
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| Co-Investigator(Kenkyū-buntansha) |
KAWAKAMI Yuki 岡山県立大学, 保健福祉学部, 助教 (30453202)
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| Co-Investigator(Renkei-kenkyūsha) |
MIYANO Masashi 青山学院大学, 理工学部, 教授 (80332287)
TOH Hiroyuki 独立行政法人産業技術総合研究所, 生命情報工学センター, 副研究センター長 (70192656)
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| Project Period (FY) |
2013-04-01 – 2016-03-31
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| Project Status |
Completed (Fiscal Year 2015)
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| Budget Amount *help |
¥5,070,000 (Direct Cost: ¥3,900,000、Indirect Cost: ¥1,170,000)
Fiscal Year 2015: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Fiscal Year 2014: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2013: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
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| Keywords | ロイコトリエン / 単鎖抗体 |
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| Outline of Final Research Achievements |
Based on the primary structure of heavy and light chains of monoclonal antibody against leukotriene C4, a key lipid mediator in chronic inflammation, we prepared a recombinant single-chain antibody comprising variable regions of these two chains and expressed it in Pichia pastoris. The affinity and recovery of the recombinant antibody was dramatically increased as compared with the antibody expressed in E. coli. The anti-LTC4 single-chain antibody as well as the original monoclonal antibody suppressed the increase in expression of mRNA for COL1A1 and COL1A2 that was induced by stimulation with LTC4 and LTD4 in fibroblasts prepared from the lung of mice. Furthermore, we exhaustively prepared mutants of the anti-LTC4 single-chain antibody based on the results of X-ray crystallography analysis of the Fab fragment of the original monoclonal antibody, and identified amino acids which determined the LT binding specificity.
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