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Diagnostic Technologies and Molecular and Cellular Profiling: Biomarker Assay Development, Validation, and Qualification

Metrology for cancer biomarkers: Affinity analysis of human HER2 and telomerase with IgYs

Biochemical Science Division, NIST, Gaithersburg, MD, Laboratory of Pathology and Urologic Oncology Branch, Center for Cancer Research, National Cancer Institute, Bethesda, MD, Division of Cancer Prevention, National Cancer Institute, Bethesda, MD

Abstract

A3

To improve quantitation of HER2 and telomerase as cancer biomarkers, we have generated novel avian polyclonal antibodies of IgY isotype⁽¹⁾ for comparison with commercially available mammalian antibodies in cell culture models. IgYs were generated against antigens for two human proteins, HER2 and telomerase, and compared with mammalian IgGs and IgMs by immunochemistry (IC), Western blotting and layered peptide array (LPA) technologies. Mammalian antibodies for both targets have been previously reported. However, metrology for quantitative comparisons of affinity reagents in general is lacking. Here we report direct comparisons between mammalian and avian reagents with several methods. These results are directed at improving metrology and validated reagents in model systems on which future clinical validation studies will be based.(1) Yan Xiao, (2006) IgY ANTIBODIES TO HUMAN TELOMERASE REVERSE TRANSCRIPTASE. Patent application has been filed (attorney docket number 000479.00163 for the US application, and filing date June 26, 2006).