Preparation of Luciferase-fused Peptides for Immunoassay of Amyloid Beta
| Autor: | Taiki Arisawa, Naomi Sakono, Takuma Ohya, Masafumi Sakono, Atsushi Manaka |
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| Rok vydání: | 2021 |
| Předmět: |
chemistry.chemical_classification
Immunoassay Amyloid Amyloid beta-Peptides biology medicine.diagnostic_test Amyloid beta Peptide Enzyme-Linked Immunosorbent Assay Molecular biology Fusion protein Antibodies Analytical Chemistry chemistry.chemical_compound chemistry Antigen Coelenterazine biology.protein medicine Luciferase Surface plasmon resonance Luciferases |
| Zdroj: | Analytical sciences : the international journal of the Japan Society for Analytical Chemistry. 37(5) |
| ISSN: | 1348-2246 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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