Preparation of Luciferase-fused Peptides for Immunoassay of Amyloid Beta

Masafumi Sakono*, Taiki Arisaw, Takuma Ohya, Naomi Sakono, Atsushi Manaka

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

1 Scopus citations

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 languageEnglish
Pages (from-to)759-763
Number of pages5
JournalAnalytical Sciences
Volume37
Issue number5
DOIs
StatePublished - 2021/05

Keywords

  • Amyloid beta
  • enzyme-labeled antigen
  • fusion protein
  • luciferase

ASJC Scopus subject areas

  • Analytical Chemistry

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