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Detection of proteins using a colorimetric bio-barcode assay

Nature Protocols volume 2, pages 1438–1444 (2007)Cite this article

Abstract

The colorimetric bio-barcode assay is a red-to-blue color change-based protein detection method with ultrahigh sensitivity. This assay is based on both the bio-barcode amplification method that allows for detecting miniscule amount of targets with attomolar sensitivity and gold nanoparticle-based colorimetric DNA detection method that allows for a simple and straightforward detection of biomolecules of interest (here we detect interleukin-2, an important biomarker (cytokine) for many immunodeficiency-related diseases and cancers). The protocol is composed of the following steps: (i) conjugation of target capture molecules and barcode DNA strands onto silica microparticles, (ii) target capture with probes, (iii) separation and release of barcode DNA strands from the separated probes, (iv) detection of released barcode DNA using DNA-modified gold nanoparticle probes and (v) red-to-blue color change analysis with a graphic software. Actual target detection and quantification steps with premade probes take ∼3 h (whole protocol including probe preparations takes ∼3 days).

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Figure 1: Colorimetric bio-barcode amplification assay.
Figure 2: DNA-modified gold nanoparticle-based barcode DNA detection and quantification.
Figure 3: Gold nanoparticle-based colorimetric barcode DNAIL-2 detection (left, barcode capture DNA-modified 30 nm gold nanoparticle spots on a TLC plate; right, quantification data).

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Acknowledgements

This work was supported by the Korea Research Foundation Grant funded by Korea Government (MOEHRD, Basic Research Promotion Fund; KRF-2006-C00217), the Ministry of Commerce and Seoul RNBD program(11001).

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Authors and Affiliations

  1. Department of Chemistry, Seoul National University, San 56-1, Sillim-dong, Seoul, 151-747, Gwanak-gu, South Korea

    Jwa-Min Nam

  2. Interdisciplinary Program in Nano-Science and Technology, Seoul National University, San 56-1, Sillim-dong, Seoul, 151-747, Gwanak-gu, South Korea

    Jwa-Min Nam & Kyung-Jin Jang

  3. Department of Chemistry, Berkeley and Physical Biosciences Division, University of California, Lawrence Berkeley National Laboratory, 94720, California, USA

    Jay T Groves

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Correspondence to Jwa-Min Nam or Jay T Groves.

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Nam, JM., Jang, KJ. & Groves, J. Detection of proteins using a colorimetric bio-barcode assay. Nat Protoc 2, 1438–1444 (2007). https://doi.org/10.1038/nprot.2007.201

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