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Whole-blood immunoassay for γH2AX as a radiation biodosimetry assay with minimal sample preparation

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Abstract

The current state of the art in high-throughput minimally invasive radiation biodosimetry involves the collection of samples in the field and analysis at a centralized facility. We have developed a simple biological immunoassay for radiation exposure that could extend this analysis out of the laboratory into the field. Such a forward placed assay would facilitate triage of a potentially exposed population. The phosphorylation and localization of the histone H2AX at double-stranded DNA breaks has already been proven to be an adequate surrogate assay for reporting DNA damage proportional to radiation dose. Here, we develop an assay for phosphorylated H2AX directed against minimally processed sample lysates. We conduct preliminary verification of H2AX phosphorylation using irradiated mouse embryo fibroblast cultures. Additional dosimetry is performed using human blood samples irradiated ex vivo. The assay reports H2AX phosphorylation in human blood samples in response to ionizing radiation over a range of 0–5 Gy in a linear fashion, without requiring filtering, enrichment, or purification of the blood sample.

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Acknowledgments

This work was supported by Grant No. U19 AI067773, the Center for High-Throughput Minimally Invasive Radiation Biodosimetry, from the National Institutes of Health/National Institute of Allergy and Infectious Diseases. This work was also supported by SBIR Phase I Grant No. 1314228 from the National Science Foundation. We thank John Seabrook for assistance in conducting some of the experiments.

Conflict of interest

The authors share the following competing interests: MLJ is a principal in Bialanx, Inc., which was previously awarded an NSF SBIR Phase I grant (#1314228). For the remaining authors, none were declared.

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    Authors and Affiliations

    1. Bialanx, Inc., 511 Avenue of the Americas, Suite 267, New York, NY, USA

      Matthew L. Johnston

    2. Center for Radiological Research, Columbia University College of Physicians and Surgeons, 630 W. 168th St., New York, NY, USA

      Erik F. Young

    3. Department of Electrical Engineering, Columbia University, 550 W 120th St., New York, NY, USA

      Matthew L. Johnston & Kenneth L. Shepard

    4. Department of Biomedical Engineering, Columbia University, 500 W 120th St., New York, NY, USA

      Kenneth L. Shepard

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    1. Matthew L. Johnston
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    2. Erik F. Young
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    Correspondence to Matthew L. Johnston.

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    Matthew L. Johnston and Erik F. Young have contributed equally to this work.

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    Johnston, M.L., Young, E.F. & Shepard, K.L. Whole-blood immunoassay for γH2AX as a radiation biodosimetry assay with minimal sample preparation. Radiat Environ Biophys 54, 365–372 (2015). https://doi.org/10.1007/s00411-015-0595-4

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    • DOI: https://doi.org/10.1007/s00411-015-0595-4

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