Voltammetric Homogeneous Binding Assay of Biotin without a Separation Step Using Iminobiotin Labeled with an Electroactive Compound

• SUGAWARA Kazuharu

  Faculty of Education, Gunma University

• KAMIYA Naoto

  Faculty of Education, Gunma University

• HIRABAYASHI George

  Faculty of Education, Gunma University

• KURAMITZ Hideki

  Department of Environmental Biology and Chemistry, Faculty of Science, Toyama University

抄録

Avidin, which is one type of glycoprotein, has a strong affinity with biotin (K_a = 10¹⁵ M^-1). Iminobiotin also forms a complex with avidin (K_a = 10⁸ M^-1 at pH 9.5). The avidin-iminobiotin complex changes to the avidin-biotin complex in the presence of biotin because of the difference of the binding constant to avidin. In this study, the interaction between avidin and iminobiotin labeled with an electroactive compound was investigated by voltammetry. After avidin and the labeled iminobiotin (LI) were incubated in 0.1 M phosphate buffer (pH 7.0), the peak currents of LI were measured in various concentrations of biotin. The peak currents increased with increasing the concentration of biotin. Thus, this observation indicates the formation of avidin-biotin complex. On the other hand, the formation of avidin-iminobiotin complex depended on the pH of the solution. LI combines with the avidin at pH 5.6 - 8.9 and dissociates at pH 4.6.

収録刊行物

• Analytical Sciences

  Analytical Sciences 21 (8), 897-900, 2005

  社団法人 日本分析化学会

キーワード

• Analytical Chemistry

詳細情報

• CRID: 1390001204258497408
• NII論文ID: 130004441113
• NII論文ID: 10016644425
• NII書誌ID: AA10500785
• DOI: 10.2116/analsci.21.897
• COI: 1:CAS:528:DC%2BD2MXoslGgt7o%3D
• ISSN: 13482246
• ISSN: 09106340
• NDL書誌ID: 7383879
• PubMed: 16122157
• Web Site: http://id.ndl.go.jp/bib/7383879
• Web Site: https://ndlsearch.ndl.go.jp/books/R000000004-I7383879
• Web Site: https://link.springer.com/content/pdf/10.2116/analsci.21.897.pdf
• Web Site: https://link.springer.com/article/10.2116/analsci.21.897/fulltext.html
• Web Site: http://www.jstage.jst.go.jp/article/analsci/21/8/21_8_897/_pdf
• 本文言語コード: en
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