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A Surface Plasmon Resonance Sensor for Substance P Using Gold-Modified Calmodulin and Melittin

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Abstract

A surface plasmon resonance (SPR) biosensor for the quantification of a neuropeptide substance P (SP) is described based on an inhibition assay using Au colloid-modified calmodulin (Au-CaM) and a target peptide melittin immobilized on carboxymethylated dextran. The modification of CaM with streptavidin Au colloids was achieved in a sample solution by the amine coupling method. The SPR signal sharply increased, corresponding to the formation of a Ca2+-Au-CaM-melittin complex on the sensor surface, and approached a steady state within 5 min. When SP was added to a sample solution, the SPR signal was decreased, due to the formation of a Ca2+-Au-CaM-SP complex in the sample solution. The modification of CaM with streptavidin Au colloids was effective for enhancing the SPR signal for SP. A decrease in the SPR signal was observed for SP in the concentration range from 0.10 to 5.0 μM, whose lower limit was ten-times superior to that (1.0 μM) with unmodified CaM. The response was highly selective to SP and the selectivity was in the order of SP ≫ neurokinin A > neurokinin B > neurotransmitters (glycine, GABA, L-glutamate, acetylcholine, norepinephrine, 5HT) ≈ substance P fragment (1–7). The potential use of the present sensor for the quantification of SP in mouse brain extracts is demonstrated.

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Authors and Affiliations

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

    Takanori Karasawa & Masao Sugawara

Authors
  1. Takanori Karasawa
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  2. Masao Sugawara
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Correspondence to Masao Sugawara.

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Karasawa, T., Sugawara, M. A Surface Plasmon Resonance Sensor for Substance P Using Gold-Modified Calmodulin and Melittin. ANAL. SCI. 21, 1431–1436 (2005). https://doi.org/10.2116/analsci.21.1431

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  • DOI: https://doi.org/10.2116/analsci.21.1431

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