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Highly Sensitive and Rapid Assay of Substance P and Streptolysin O in Human Serum Using Immuno-liposomes and Gramicidin Channels

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Abstract

A highly sensitive and rapid method for the determination of substance P (SP) and streptolysin O (SLO) in human serum is described. The assay is based on enriching the analyte by agglutination/precipitation of immuno-liposomes and enhancing the fluorescence intensity by gramicidin A channels. A mixture of the immuno-liposomes encapsulating a pH-sensitive fluorescence dye BCECF, gramicidin A and a given concentration of SP (or SLO) is preincubated in a solution and captured on anti-SP (or anti-SLO)-modified cover slips, followed by measuring fluorescence images after removing excess liposomes. The method allowed quantifying SP and SLO in the range from sub-pg mL−1 to pg mL−1, with detection limits of 0.32 pg mL−1 and 8 fg mL−1, respectively. The present method could determine SP and SLO in 50 - 125 times diluted human serum without any extraction steps. The assay can be completed within 60 min.

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Authors and Affiliations

  1. Department of Chemistry, College of Humanities and Sciences, Nihon University, Sakurajyosui, Setagaya, Tokyo, 156-8550, Japan

    Misato Sakamoto & Masao Sugawara

  2. School of Pharmacy, Tokyo University of Pharmacy and Life Science, 1432-1 Horinouchi, Hachioji, Tokyo, 192-0392, Japan

    Atsushi Shoji

Authors
  1. Misato Sakamoto
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  2. Atsushi Shoji
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  3. Masao Sugawara
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Correspondence to Masao Sugawara.

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Sakamoto, M., Shoji, A. & Sugawara, M. Highly Sensitive and Rapid Assay of Substance P and Streptolysin O in Human Serum Using Immuno-liposomes and Gramicidin Channels. ANAL. SCI. 29, 877–883 (2013). https://doi.org/10.2116/analsci.29.877

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  • DOI: https://doi.org/10.2116/analsci.29.877

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