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The use of camel antibodies in development of EGFRvIII enzyme-linked immunosorbent assay

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Abstract

In the present study, anti-EGFRvIII antibodies were purified from the serum of an immunized Arabian camel (Camelus dromedarius). The three subclasses of IgG were purified using protein-G column chromatography. Epidermal growth factor receptor vIII (EGFRvIII) was purified from HC220d2/c cell line expressed the highest levels of EGF receptors using a concanavalin A column chromatography. The ELISA procedure was developed using 2 types of enzyme labels namely horse radish peroxidase (HRP) and penicillinase. The affinity constants of purified total IgG and Ig fractions (IgG1, IgG2 and IgG3) calculated were found to be 4.48 × 107, 1.3 × 107, 4.23 × 106 and 1.67 × 106 M−1, respectively. The sensitivity of assays for the detection of EGFR when used total IgG were found to be 0.35 μg/mL in HRP label assay and 0.45 μg/mL in penicillinase label assay. We concluded that the ELISA in competitive or inhibition manners using polyclonal camel antibodies containing total IgG (but not IgG1, IgG2, and IgG3 fractions) can be promising in detection of EGFRvIII in biological fluids.

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Authors and Affiliations

  1. Department of Clinical Biochemistry, School of Medical Sciences, Tarbiat Modares University, Tehran, Iran

    A. Naderi Sohi & M. J. Rasaee

  2. Department of Clinical Biochemistry, Faculty of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran

    M. Rajabibazl

  3. School of Advanced Technologies in Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran

    M. Rajabibazl

  4. Endocrinology and Metabolism Research Center, Tehran University of Medical Sciences, Tehran, Iran

    K. Omidfar

Authors
  1. A. Naderi Sohi
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  2. M. Rajabibazl
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  3. M. J. Rasaee
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  4. K. Omidfar
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Correspondence to M. Rajabibazl.

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Sohi, A.N., Rajabibazl, M., Rasaee, M.J. et al. The use of camel antibodies in development of EGFRvIII enzyme-linked immunosorbent assay. Appl Biochem Microbiol 51, 374–380 (2015). https://doi.org/10.1134/S0003683815030163

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  • DOI: https://doi.org/10.1134/S0003683815030163

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