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

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.