Analytical Sciences

Preparation of Luciferase-fused Peptides for Immunoassay of Amyloid Beta

Masafumi SAKONO,* Taiki ARISAWA,* Takuma OHYA,* Naomi SAKONO,** and Atsushi MANAKA**

*Department of Applied Chemistry, Faculty of Engineering, University of Toyama, 3190 Gofuku, Toyama, Toyama 930-8555, Japan
**Department of Applied Chemistry and Chemical Engineering, National Institute of Technology, Toyama College, 13 Hongo, Toyama, Toyama 393-8630, Japan

An immunoassay, such as the enzyme-linked immunosorbent assay (ELISA), is an analytical method that utilizes the interaction of antigens and antibodies. Enzyme-labeled antigens require both molecular recognition by the antibody and enzymatic activity as a reporter. We designed and constructed an immunodetection system for amyloid beta peptides (Aβ) using an enzyme-labeled antigen expressed from Escherichia coli. Aβ(1–16) fused with renilla luciferase was prepared as the enzyme-labeled antigen. In the presence of this luciferase-fused peptide, the luminescence of coelenterazine-h was observed. The influence of the fusion with Aβ on the luminescence reaction was insignificant. Surface plasmon resonance analysis indicated that the interaction between the luciferase-fused Aβ and anti-Aβ antibody was sufficiently strong. In the competitive ELISA assay for Aβ detection using the luciferase-fused Aβ, the luminescence intensity decreased as the Aβ concentration increased.

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