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SNAP-Tag Technology: A Promising Tool for Ex Vivo Immunophenotyping

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Abstract

Background

SNAP-tag, a self-labeling protein tag, is commonly used for in vitro and in vivo analysis of bound target proteins. We report the first evidence that SNAP-tag could be used for ex vivo detection of enriched biological markers.

Methods

Proof of concept was established for target c-kit receptor, a pathological and diagnostic marker for a variety of cancers. SNAP-tag conjugates with stem-cell factor (SCF) fusion proteins were designed and their binding and specificity was validated in vitro using flow cytometry and immunostaining.

Results

Ex vivo diagnostic application of the fusion protein was demonstrated in comparison with anti-c-kit antibody for peripheral blood samples from leukemia patients and colorectal tissue specimens.

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Acknowledgements

We would like to thank Ahmad Fawzi Hussain, Alessa Pardo and Reinhard Rosinke (Department of Pharmaceutical Product Development, Fraunhofer Institute for Molecular Biology and Applied Ecology) for their assistance with labeling experiments and protein purification, respectively, and DAAD and MHRD for providing fellowships to SC.

Author information

Author notes

  1. Stefan Barth

    Present address: UCT Faculty of Health Sciences, South African Research Chair in Cancer Biotechnology, Cape Town, South Africa

Authors and Affiliations

  1. Department of Biotechnology, Bhupat and Jyoti Mehta School of Biosciences, Indian Institute of Technology Madras, Chennai, 600036, India

    Swati Choudhary & Rama S. Verma

  2. Department of Pharmaceutical Product Development, Fraunhofer Institute for Molecular Biology and Applied Ecology IME, 52074, Aachen, Germany

    Stefan Barth

Authors
  1. Swati Choudhary
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  2. Stefan Barth
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Corresponding authors

Correspondence to Stefan Barth or Rama S. Verma.

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Conflict of interest

SC, SB and RSV declare that they have no conflict of interest.

Funding

This work was funded by the block grants at Indian Institute of Technology Madras and Fraunhofer IME.

Ethical approval

All procedures performed in studies involving human participants were in accordance with The Code of Ethics of the World Medical Association (Declaration of Helsinki) for experiments involving humans and were approved by the Indian Institute of technology Madras Institutional Review Board (IEC/2016/01/RSV-6/22). Informed consent was obtained from all individual participants included in the study.

Additional information

S. Barth and R. S. Verma contributed equally to this work.

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Choudhary, S., Barth, S. & Verma, R.S. SNAP-Tag Technology: A Promising Tool for Ex Vivo Immunophenotyping. Mol Diagn Ther 21, 315–326 (2017). https://doi.org/10.1007/s40291-017-0263-2

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  • DOI: https://doi.org/10.1007/s40291-017-0263-2

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