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An application of capsid-specific artificial ankyrin repeat proteinproduced in E. coli for immunochromatographic assay as a surrogate for antibody

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Abstract

Immunochromatographic strip test is a unique type of rapid test that has been developed for use as part of a diagnostic kit for the rapid detection of antibodies and/or other proteins of interest. For the detection of target proteins, most of the commercial tests are assembled based on the conjugation of colloidal gold particles to monoclonal antibodies embedded within the conjugate pad of a strip test. In this study, we tested the novel concept of using an artificial non-antibody structure for generating a colloidal gold conjugate (CGC). We exploited the property of an ankyrin repeat protein that specifically binds to the HIV-1 capsid protein termed AnkGAG1D4. This construct was applied as a model structure to create Ank1D4-CGC and used as a new type of visible detector system and termed it ankyrin-based immunochromatographic strip (ABIS) test. The ABIS test was shown to be highly sensitive with a lower limit of detection of the target protein at 0.1 μg/ml. Moreover, the ABIS test was not only highly sensitive but also shared a level of specificity within the same range of the commercial test kit. The results of the studies presented herein therefore demonstrate the novel application of an artificial non-immunoglobulin structure (ankyrin repeat protein) as the new line of a visible detector using a rapid diagnostic test with characteristics that have the potential to be superior to those that utilize antibody-based tests.

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Acknowledgments

The work in Thailand was supported by the National Research University project under the Thailand’s Office of the Commission on Higher Education, the Research Chair Grant of National Sciences and the Technology Development Agency at the Faculty of Associated Medical Sciences, Chiang Mai University, School of Allied Health Sciences, University of Phayao, Thailand Research Fund (TRF), National Research Council of Thailand (NRTC), Health Systems Research Institute (HSRI), and i + Med Laboratories Company Limited.

Conflict of interest

None.

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Authors and Affiliations

  1. Division of Clinical Immunology and Transfusion Sciences, School of Allied Health Sciences, University of Phayao, Phayao, 56000, Thailand

    Sawitree Nangola

  2. Department of Pathology, Emory University School of Medicine, Atlanta, GA, 30332, USA

    Aftab A. Ansari

  3. Division of Clinical Immunology, Faculty of Associated Medical Sciences, Chiang Mai University, Chiang Mai, 52000, Thailand

    Weeraya Thongkum & Somphot Saoin

  4. Division of Clinical Immunology and Biomedical Technology Research Unit, National Center for Genetic Engineering and Biotechnology, National Science and Technology Development Agency Faculty of Associated Medical Sciences, Chiang Mai University, Chiang Mai, 52000, Thailand

    Chatchai Tayapiwatana

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  1. Sawitree Nangola
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  2. Weeraya Thongkum
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  3. Somphot Saoin
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  4. Aftab A. Ansari
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  5. Chatchai Tayapiwatana
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Correspondence to Chatchai Tayapiwatana.

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Nangola, S., Thongkum, W., Saoin, S. et al. An application of capsid-specific artificial ankyrin repeat proteinproduced in E. coli for immunochromatographic assay as a surrogate for antibody. Appl Microbiol Biotechnol 98, 6095–6103 (2014). https://doi.org/10.1007/s00253-014-5777-5

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  • DOI: https://doi.org/10.1007/s00253-014-5777-5

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