ELISABioassayQualityChecker

ELISA Bioassay Quality Checker — Control of ELISA Quality

ℹ️  Utility checks critical parameters of ELISA validity:
   • Calibration Curve Linearity (R²): ≥0.99
   • Standard Recovery: 80–120%
   • Duplicate Precision (CV): ≤15%
   • Blank Optical Density (Blank OD): <0.1 (example)

⚠️  CRITICAL: If any parameter fails criteria, the entire run is considered invalid!
   This is a GLP/GMP requirement for bioanalytical methods.

Usage:
 ELISABioassayQualityChecker.exe                            → demo mode (console output)
 ELISABioassayQualityChecker.exe input.csv output.json      → evaluate your data

Input format:
BatchNumber,CurveRSquared,StandardRecoveryPercent,DuplicateCVPercent,BlankOD,CalculatedConcentration

Example:
 ELISA-BIO-2026-001,0.998,98.5,4.2,0.05,125.5

— WHY IS THIS NEEDED?
ELISA is widely used for quantification of proteins, antibodies, vaccines, and biomarkers.
• Assay quality depends on many factors: incubation, washing, conjugate activity.
• Calibration curve must be linear (or log-linear) with high R².
• Recovery indicates method accuracy (absence of matrix effects).
• Duplicate CV reflects pipetting precision and reaction homogeneity.
• High Blank may indicate non-specific binding or contamination.
• Automation of criteria check eliminates operator subjectivity.

⚠️  CRITICAL:
• R² < 0.99 often requires curve recalculation or standard re-dilution.
• CV > 15% indicates technical error (bubbles, uneven washing).
• Recovery outside 80-120% may require buffer or extraction changes.
• All criteria must be pre-approved in the method validation protocol.

Key features:
• Comprehensive check of all main ELISA QC parameters.
• Support for microplate reader data (Molecular Devices, Tecan, BioTek).
• Generation of run validity report.

Critical parameters:
• Curve R²: >= 0.99
• Recovery: 80-120%
• Duplicate CV: <= 15%
• Blank OD: < Limit

💡 Usage tips:
1. Use 4-parameter logistic regression (4PL) for curve fitting if sigmoidal.
2. Always include Blank and Zero Standard.
3. Check plate edges for evaporation effect (edge effect).
4. Use control samples (QC Low, Mid, High) in each run.
5. Regularly calibrate reader and check filters.

⚠️ Note: Unlike chemical analysis, bioassays have higher variability. Therefore, acceptance criteria (especially CV) may be wider (up to 20-25% at the Lower Limit of Quantitation LLOQ).