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Recombinant human chymase produced by silkworm-baculovirus expression system: its application for a chymase detection
Takeo Suzuki1, Hiroki Kaki, Shinichi Naya
1Chuo-Sanken Laboratory, Katakura Industries Co., Ltd., Saitama, Japan. tk.suzuki@katakura.co.jp
Japanese Journal of Pharmacology
|December 25, 2002
Summary
Human chymase, a mast cell enzyme, was produced recombinantly for functional studies. Elevated chymase levels were detected in hypertensive patients using a novel ELISA system.
Area of Science:
- Biochemistry
- Enzymology
- Immunology
Background:
- Human chymase is a mast cell-derived serine proteinase involved in various diseases.
- Its precise physiological and pathophysiological roles in vivo remain unclear.
Purpose of the Study:
- To produce large quantities of recombinant human chymase.
- To develop an ELISA system for measuring chymase concentration.
- To investigate chymase's role in hypertension.
Main Methods:
- Recombinant human chymase was produced using a silkworm-baculovirus expression system.
- The enzyme was purified and characterized.
- Anti-chymase monoclonal antibodies were generated to develop an ELISA system.
Main Results:
- Recombinant human chymase was successfully produced, purified, and found to be enzymatically identical to the native enzyme.
- The recombinant enzyme exhibited high stability in cell culture media.
- Preliminary studies using the ELISA system revealed significantly higher chymase concentrations in hypertensive patients compared to normal individuals.
Conclusions:
- The developed recombinant human chymase and ELISA system are suitable for in vitro and in vivo assays.
- The findings suggest a potential role for chymase in hypertension.
- The ELISA system holds promise for clinical diagnosis and evaluating chymase-targeting drugs.