R-based workflow for quantitative analysis of hyaluronan (HA) ELISA data using four-parameter logistic (4PL) standard-curve modeling.
This project was developed to analyze hyaluronan concentrations in mouse skin and plasma samples while providing greater control over data curation, quality assessment, and reporting than a standard automated analysis workflow.
ELISA standard curves are modeled using a four-parameter logistic (4PL) regression, and the fitted curve is used to estimate HA concentrations in experimental samples.
The workflow provides results comparable to those obtained using MyAssays 4PL analysis, while allowing additional curation and inspection of assay results within R.
The analysis includes:
- Import and organization of ELISA plate data
- Identification and processing of standards and experimental samples
- 4PL standard-curve fitting
- Sample concentration estimation from the fitted curve
- Dilution-factor handling
- Quality assessment and result curation
- Visualization of standard curves and assay results
- Export of processed results for downstream analysis
The workflow is used to quantify hyaluronan levels in mouse skin and plasma samples.
Hyaluronan concentrations are estimated from ELISA measurements using the fitted 4PL standard curve, allowing experimental samples to be evaluated alongside assay standards and quality-control criteria.
Commercial ELISA analysis platforms can perform 4PL standard-curve fitting, but repeated use may require paid access or incur analysis costs.
This workflow was developed as a reproducible, no-cost alternative for routine HA ELISA analysis, while also providing greater control over data curation, quality assessment, visualization, and reporting.
The R-based implementation allows:
- Reproducible 4PL standard-curve fitting
- Sample concentration estimation without reliance on paid analysis software
- Direct inspection of fitted curves and calculated concentrations
- Greater control over inclusion and exclusion of assay results
- Customized quality assessment and data curation
- Flexible visualization and reporting
- Reanalysis of historical data using the same computational workflow
Results can be compared with those generated using commercial 4PL analysis tools such as MyAssays, while the underlying analysis remains transparent and reproducible within R.
Language: R
Methods: ELISA · four-parameter logistic (4PL) regression · standard-curve modeling · concentration estimation · assay quality assessment · data visualization