Production and Application of Monoclonal Antibodies to Human Selenoprotein P
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概要
- 論文の詳細を見る
Selenoprotein P is a selenium-rich extracellular protein and is the major selenoprotein in plasma. Although the biological function of selenoprotein P has not been established, we have recently shown that selenoprotein P has phospholipid hydroperoxide glutathione peroxidase-like activity. To study the structure and function of selenoprotein P, we produced monoclonal antibodies against human selenoprotein P. Immunization of rats with purified selenoprotein P was followed by hybridization, cloning, and the establishment of eleven hybridomas producing specific antibodies against human selenoprotein P. With the addition of six kinds of insoluble rat anti-human selenoprotein P monoclonal antibodies to human plasma, the selenium concentration of the supernatant was decreased to 47% of the control. This suggests that selenoprotein P constituted 53% of total plasma selenium. Western blot analysis of the immunoprecipitate from human plasma showed that the antibodies specifically bound to a 69 kDa protein, representing selenoprotein P. Next, we developed an enzyme-linked immunosorbent assay for selenoprotein P using two monoclonal antibodies. The plasma concentration of selenoprotein P in 77 normal individuals was 5.3 ± 1.1 μg/ml. Because preferential depletion of selenoprotein P by low density lipoprotein(LDL)-apheresis has been reported, we next measured selenoprotein P concentration of plasma samples from patients before and after LDL-apheresis. We confirmed that the plasma concentration of selenoprotein P was decreased from 5.7 to 2.3 μg/ml by LDL-apheresis. This result shows that this assay provides a reliable means of measuring the content of selenoprotein P in plasma.
- 社団法人日本薬学会の論文
- 2001-08-01
著者
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HONJOH Tsutomu
Morinaga Institute of Biological Science, Inc.
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Honjoh Tsutomu
Morinaga Institute Of Biological Science Inc.
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Takahashi Kazuhiko
Department of Drug Metabolism and Disposition, Graduate School of Pharmaceutical Sciences, Nagoya Ci
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Saito Yoshiro
Department Of Hygienic Chemistry Graduate School Of Pharmaceutical Sciences Hokkaido University
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Watanabe Yasuko
Department of Hygienic Chemistry, Graduate School of Pharmaceutical Sciences, Hokkaido University
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Saito Eiji
Second Department of Internal Medicine, Nihon University School of Medicine
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Watanabe Yasuko
Department Of Hygienic Chemistry Graduate School Of Pharmaceutical Sciences Hokkaido University
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Honjoh T
Morinaga Institute Of Biological Sciences
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Saito Eiji
Second Department Of Internal Medicine Nihon University School Of Medicine
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Takahashi Kazuhiko
Department Of Cardiovascular Diseases Miyagi Prefectural Semine Hospital
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Takahashi Kazuhiko
Department Of Applied Chemistry Tokyo Institute Of Technology
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Takahashi Kazuhiko
Department Of Hygienic Chemistry Graduate School Of Pharmaceutical Sciences Hokkaido University
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Saito Yoshihiro
Department Of Aeronautics And Astronautics
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HONJOH Tsutomu
Morinaga Institute of Biological Science
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