Abstract
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.
Original language | English |
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Pages (from-to) | 759-763 |
Number of pages | 5 |
Journal | Analytical Sciences |
Volume | 37 |
Issue number | 5 |
DOIs | |
State | Published - 2021/05 |
Keywords
- Amyloid beta
- enzyme-labeled antigen
- fusion protein
- luciferase
ASJC Scopus subject areas
- Analytical Chemistry