Enzyme-linked immunosorbent assay for glycyrrhizin using anti-glycyrrhizin monoclonal antibody and an eastern blotting technique for glucuronides of glycyrrhetic acid

S. Shan, Hiroyuki Tanaka, Y. Shoyama

研究成果: ジャーナルへの寄稿記事

81 引用 (Scopus)

抄録

Hybridomas secreting a monoclonal antibody against glycyrrhizin were produced by fusing splenocytes from a mouse immunized against a glycyrrhizin - bovine serum albumin conjugate with the hypoxanthine - aminopterin - thymidine-sensitive mouse myeloma cell line, P3-X63-Ag8-653. A very weak cross-reaction with glycyrrhetinic acid monoglucuronide and glycyrrhetic acid occurred. An enzyme-linked immunosorbent assay (ELISA) that had an effective measuring range of 20-200 ng/mL of glycyrrhizin was established using this monoclonal antibody. In addition, a method named eastern blotting for the detection of glycyrrhizin was investigated. In this method, we developed a new way to separate the glycyrrhizin molecule into two functional parts using a simple and well-known chemical reaction. The sugar parts were oxidized by sodium periodate to give dialdehydes, which reacted with amino groups on the protein and covalently bound to the adsorbent membrane. The monoclonal antibody bound to the aglycone part of the glycyrrhizin molecule for immunostaining. This method was validated by immunocytolocalization of glycyrrhizin in fresh Glycyrrhiza root.

元の言語英語
ページ(範囲)5784-5790
ページ数7
ジャーナルAnalytical Chemistry
73
発行部数24
DOI
出版物ステータス出版済み - 12 15 2001

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Glycyrrhetinic Acid
Glycyrrhizic Acid
Immunosorbents
Glucuronides
Assays
Monoclonal Antibodies
Enzymes
Aminopterin
Molecules
Hypoxanthine
Bovine Serum Albumin
Sugars
Thymidine
Adsorbents
Chemical reactions
Cells
Membranes

All Science Journal Classification (ASJC) codes

  • Analytical Chemistry

これを引用

Enzyme-linked immunosorbent assay for glycyrrhizin using anti-glycyrrhizin monoclonal antibody and an eastern blotting technique for glucuronides of glycyrrhetic acid. / Shan, S.; Tanaka, Hiroyuki; Shoyama, Y.

:: Analytical Chemistry, 巻 73, 番号 24, 15.12.2001, p. 5784-5790.

研究成果: ジャーナルへの寄稿記事

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abstract = "Hybridomas secreting a monoclonal antibody against glycyrrhizin were produced by fusing splenocytes from a mouse immunized against a glycyrrhizin - bovine serum albumin conjugate with the hypoxanthine - aminopterin - thymidine-sensitive mouse myeloma cell line, P3-X63-Ag8-653. A very weak cross-reaction with glycyrrhetinic acid monoglucuronide and glycyrrhetic acid occurred. An enzyme-linked immunosorbent assay (ELISA) that had an effective measuring range of 20-200 ng/mL of glycyrrhizin was established using this monoclonal antibody. In addition, a method named eastern blotting for the detection of glycyrrhizin was investigated. In this method, we developed a new way to separate the glycyrrhizin molecule into two functional parts using a simple and well-known chemical reaction. The sugar parts were oxidized by sodium periodate to give dialdehydes, which reacted with amino groups on the protein and covalently bound to the adsorbent membrane. The monoclonal antibody bound to the aglycone part of the glycyrrhizin molecule for immunostaining. This method was validated by immunocytolocalization of glycyrrhizin in fresh Glycyrrhiza root.",
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