Immunosorbent assay using gold colloid cluster technology for determination of IgEs in patients’ sera

Abstract

This study focuses on the development of a sensitive and simple cluster-linked immunosorbent assay (CLISA) using gold colloidal cluster labeling for determination of proteins such as antigens (Ags) or antibodies (Abs). Abs for detection can be labeled with gold colloid clusters (GCCs). The Fc domain of the Abs binds to the clusters, and the Fab domain to the Ag on a nitrocellulose membrane or a microtiter plate as a support for dot-blotting. The signal of positive interaction between GCC-labeled Abs and its dotted Ag is detectable by the naked eye and can be quantified by comparison to a color scale prepared from a dilution series of known sample concentrations. The colored reaction product is stable for prolonged periods and does not fade, making this method a simple, fast, and convenient means for detection of Ag or Ab biorecognitions and an alternative to enzyme-linked immunosorbent assay. Several interactions between different Ags or Abs (eg, β-lactoglobulin) and solutions avoiding gold colloidal cluster flocculation (eg, using protein G) were studied. CLISA was tested for other analytical purposes such as detection of IgEs in patients’ sera.

Keywords:

• ELISA
• allergen
• patient sera
• CLISA
• immunoassay
• β-lactoglobulin

Acknowledgments

The authors thank the Department of Biochemistry, University of Vienna (Dr Bohr Gasse 9, A-1030 Vienna, Austria) for the financial support and Daniela Gruber (Institute of Cell Imaging and Ultrastructure Research, Althanstrasse 14 A-1091 Vienna, Austria) for TEM images.

Disclosure

The authors report no conflicts of interest in this work.