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Development and Validation of an Ultrasensitive Single Molecule Array Digital Enzyme-linked Immunosorbent Assay for Human Interferon-α
Published on: June 14, 2018
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Dual amplification enabled counting based ultrasensitive enzyme-linked immunosorbent assay.
Haomin Liu1, Yu Lei2
1Department of Chemical and Biomolecular Engineering, University of Connecticut, Storrs, CT, 06269, USA.
Analytica Chimica Acta
|February 22, 2022
Summary
A novel dual amplification ELISA method enhances sensitivity for detecting low-abundance biomarkers. This technique significantly improves the detection limit for mouse IgG, offering a new approach for ultrasensitive biomarker analysis.
Area of Science:
- Biochemistry
- Analytical Chemistry
- Biotechnology
Background:
- Enzyme-linked immunosorbent assay (ELISA) is a primary method for biomarker detection.
- Detecting low-abundance biomarkers with ELISA requires enhanced sensitivity.
Purpose of the Study:
- To develop an ultrasensitive ELISA technique for detecting low-abundance biomarkers.
- To improve the sensitivity of conventional plate-based ELISA.
Main Methods:
- Developed a dual amplification strategy using tyramide signal amplification (TSA) and alkaline phosphatase.
- Enabled fluorescent precipitate formation for counting-based detection.
- Applied the method to detect mouse total IgG as a model biomarker.
Main Results:
- Achieved a lower limit of detection of 54.5 pg/mL for mouse IgG, a significant improvement from the commercial kit's 1.56 ng/mL.
- Demonstrated good linearity between precipitate count and IgG concentration.
- Recovery tests confirmed the reliability of the developed ultrasensitive ELISA method.
Conclusions:
- The dual amplification ELISA provides ultrasensitive detection of biomarkers.
- This method enhances the sensitivity of conventional plate-based ELISA, opening new avenues for biomarker analysis.
