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Fluorogenic enzyme-linked immunosorbent assay with a dual color variation
1Department of Biological and Chemical Engineering, Hongik University, 2639 Sejong-ro, Sejong-si 30016, South Korea.
Insights
A new fluorogenic enzyme-linked immunosorbent assay (FELISA) enables sensitive detection of tumor necrosis factor alpha (TNFα) and programmed cell death-ligand 1 (PDL1). This high-throughput method offers rapid and accurate biomarker analysis for disease diagnosis and treatment.
Area of Science:
- Biochemistry
- Immunology
- Assay Development
Background:
- Accurate biomarker detection is vital for disease diagnosis, monitoring, and treatment.
- Existing immunoassay tools may lack the required sensitivity or throughput for certain applications.
Purpose of the Study:
- To develop a sensitive and high-throughput fluorogenic enzyme-linked immunosorbent assay (FELISA).
- To detect key biomarkers: tumor necrosis factor alpha (TNFα) and programmed cell death-ligand 1 (PDL1).
Main Methods:
- Development of a FELISA utilizing Amplex Red or QuantaBlu fluorescent substrates.
- Application of the assay for the detection of TNFα and PDL1.
Main Results:
- Achieved a limit of detection in the nanogram order for TNFα and PDL1.
- Enabled convenient assaying of multiple samples within 20 hours.
- Demonstrated FELISA's applicability as a powerful immunoassay tool.
Conclusions:
- FELISA provides a rapid and accurate method for detecting TNFα and PDL1.
- This assay platform is versatile and can be adapted for various fluorogenic immunoassays against other antigens.
Abstract:
The development of accurate and high-throughput biomarker detection tools is crucial for the diagnosis, monitoring, and treatment of various diseases. In this study, a sensitive fluorogenic enzyme-linked immunosorbent assay (FELISA) using Amplex Red or QuantaBlu fluorescent substrate was developed for the detection of tumor necrosis factor alpha and programmed cell death-ligand 1. The limit of detection of FELISA was in the nanogram order and multiple samples were conveniently assayed within 20 h using FELISA, demonstrating its applicability as a powerful immunoassay tool. FELISA can be widely used for rapid and accurate TNFα and PDL1 detection and applied to various fluorogenic immunoassays against other antigens of interest.
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