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Variability in Microplate Surface Properties and Its Impact on ELISA.
Shera Lilyanna1, Enoch Ming Wei Ng2, Shiho Moriguchi3
1Cardiovascular Research Institute, Department of Medicine, Yong Loo Lin School of Medicine, National University of Singapore, National University Health System, Singapore.
The Journal of Applied Laboratory Medicine
|February 27, 2021
Summary
Microplate surface characteristics significantly impact immunoassay variability. Gamma-irradiation treatment improves microplate surface properties, leading to more consistent assay results and reducing false positives/negatives in biomarker detection.
Area of Science:
- Biochemistry
- Materials Science
- Analytical Chemistry
Background:
- Microplate-based immunoassays are crucial for biomarker measurement in clinical and research settings.
- Assay variability from microplates can lead to inaccurate results.
- The role of microplate surfaces in assay variability is not well understood.
Purpose of the Study:
- To investigate the impact of microplate surface characteristics on immunoassay variability.
- To identify how microplate manufacturing and treatment affect assay performance.
Main Methods:
- Used a validated sandwich ELISA to measure well-to-well assay variation in treated and untreated microplates.
- Analyzed microplate surface properties using contact angle measurements, SEM, EDX, and AFM.
Main Results:
- Observed vendor-specific assay responses and identified "rogue" plates with high variation.
- Found location-dependent bias within plates and variability in surface wettability.
- Demonstrated that gamma-irradiation alters surface properties, increasing polarity and roughness, which impacts assay responses.
Conclusions:
- Microplate surface properties like roughness, wettability, and chemistry are critical for assay consistency.
- Tighter manufacturing controls for microplates are needed to improve ELISA assay reliability.
- Addressing residual contaminants and surface characteristics can enhance immunoassay accuracy.
