Recombinant Human Chymase Produced by Silkworm-Baculovirus Expression System : Its Application for a Chymase Detection Kit
スポンサーリンク
概要
- 論文の詳細を見る
Human chymase is a mast cell-derived serine proteinase, which is a non-angiotensin converting enzyme angiotensin II-generating enzyme. It appears to participate in various diseases, but it is unclear whether chymase plays major roles in physiological and pathophysiological functions in vivo. To obtain information on the physiological and pathophysiological functions of chymase and to search for diseases in which chymase participates, in the present study, we aimed at producing recombinant human chymase in large quantities and at developing an ELISA system using anti-human chymase antibodies. A recombinant human chymase was produced by a silkworm-baculovirus expression system. The recombinant chymase in active form was efficiently purified from larval hemolymph using cation-exchange and heparin column chromatography. This recombinant enzyme was enzymatically identical with native human chymase. On the other hand, the stability of the recombinant enzyme in cultured medium for mammalian cells at 37°C was very high as compared with the stability of the native enzyme; 20% of the activity was maintained 120 h after addition of medium. These results indicated that the recombinant enzyme could also utilize in vitro and in vivo assay systems. We obtained several anti-chymase monoclonal antibodies by using the recombinant human chymase as antigen. These antibodies were used to construct an ELISA system for measuring the chymase concentration in blood. As a result of preliminary examination using this ELISA system, it was shown that the chymase concentration in each serum from hypertensive patients is significantly higher than in normal serum. The ELISA system will be applicable for clinical diagnosis and in vivo evaluation systems for chymase-targeting drugs.
- 2002-11-01
著者
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Takai Shinji
Department Of Cardiology Kouseikai Takeda Hospital
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MIYAZAKI Mizuo
Department of Pharmacology, Osaka Medical College
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Takai S
Department Of Pharmacology Osaka Medical College
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Takai Shinji
Department Of Pharmacology Osaka Medical College
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Miyazaki M
Department Of Pharmacology Osaka Medical College
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Miyazaki Mizuo
Department Of Pharmacolory Osaka City University Medical School
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NAGAI Akihiko
Fukushima Research Laboratories, TOA EIYO Ltd.,
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Nagai Akihiko
Fukushima Research Laboratories Toa Eiyo Ltd
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SUZUKI Takeo
Chuo-Sanken Laboratory, Katakura Industries Co., Ltd.
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KAKI Hiroki
Chuo-Sanken Laboratory, Katakura Industries Co., Ltd.
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NAYA Shinichi
Chuo-Sanken Laboratory, Katakura Industries Co., Ltd.
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MURAYAMA Soji
Chuo-Sanken Laboratory, Katakura Industries Co., Ltd.
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TATSUI Akira
Fukushima Research Laboratories, Toa Eiyo, Ltd
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Miyazaki Mizuo
Department Of Pharmacoldgy Osaka Medical College
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Kaki Hiroki
Chuo-sanken Laboratory Katakura Industries Co. Ltd.
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Tatsui Akira
Fukushima Research Laboratories Toa Eiyo Ltd
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Naya Shinichi
Chuo-sanken Laboratory Katakura Industries Co. Ltd.
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Murayama Soji
Chuo-sanken Laboratory Katakura Industries Co. Ltd.
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Takai Shinji
Department Of Animal Hygiene School Of Veterinary Medicine And Animal Sciences Kitasato University
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Suzuki Takeo
Chuo-sanken Laboratory Katakura Industries Co. Ltd.
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