Purpose

Define user requirements for a standalone FUZKK utility that accepts laboratory CSV data, evaluates the records using limits embedded in code, and produces LabWare-compatible JSON.

Scope

The utility is intended for preliminary QC/QA review, integration testing, LIMS/LabWare flow and evidence-trail preparation. Final release decisions remain under the laboratory's validated procedure and responsible personnel.

Users

QC analyst, QA reviewer, CSV/validation engineer, LIMS/LabWare integration engineer, responsible laboratory specialist.

User requirements

1. The utility shall run without arguments and print its self-description, a built-in input.csv example from GetDemoData(), and demo evaluation for the embedded records.
2. The utility shall run with two arguments: input.csv output.json.
3. The utility shall not read input.csv and shall not write output.json when started without arguments.
4. CSV numeric values shall be parsed using CultureInfo.InvariantCulture.
5. Output shall be generated as LabWare-compatible JSON with Header, Samples, Results, Status, StatusCode, ErrorMessage, Description and DescriptionEN.
6. For PASS records, ErrorMessage shall be an empty string.
7. Embedded limits shall follow this priority: Ph. Eur. → British Pharmacopoeia / UK implementation → EAEU / regional requirements → EMA/ICH/EU guidance → USP fallback.
8. If an exact monograph is not known, strict standard API limits are used where applicable: assay 98–102%, total impurities ≤1.0%, individual impurity ≤0.5%.
9. For biologics and mAb-like products, aggregation, sterility and endotoxin checks shall be included where relevant to the utility purpose.
10. If a parameter may arrive in different units, the unit shall be represented as a separate input field or explicitly reflected in the input.csv field name.

Input CSV

BatchNumber,ProductName,EC50_pM,Emax_Percent,HillSlope,R_Squared,RelativePotency_Percent,Z_Factor,CV_Percent,StandardEC50_pM
GLP1-BIO-2026-001,Semaglutide API,15.5,98.0,1.1,0.995,102.0,0.75,5.2,15.2
GLP1-BIO-2026-002,Liraglutide Bulk,250.0,95.0,1.0,0.990,98.0,0.70,6.5,245.0
GLP1-BIO-2026-003,Tirzepatide API,8.2,99.0,1.2,0.998,105.0,0.80,4.0,8.0

input.csv fields

Utility description

GLP-1 Potency Bioassay Reviewer — GLP-1 Biological Potency Reviewer

GLP-1 Potency Bioassay Reviewer — GLP-1 Biological Potency Reviewer

ℹ️ Utility analyzes cell-based GLP-1 activity assay data:
• Relative potency (80-125%)
• EC50 (half-maximal effective concentration)
• Emax (maximum effect)
• Curve fit quality (R²)
• Z-Factor (assay quality)
• CV (variability)

⚠️ CRITICAL: Chemical purity does not guarantee biological activity!
Deamidation or aggregation may reduce receptor binding.

Usage:
GLP1PotencyBioassayReviewer.exe → demo mode (console output)
GLP1PotencyBioassayReviewer.exe input.csv output.json → evaluate your data

Input format:
BatchNumber,ProductName,EC50_pM,Emax_Percent,HillSlope,R_Squared,RelativePotency_Percent,Z_Factor,CV_Percent,StandardEC50_pM

Example:
GLP1-BIO-2026-001,Semaglutide API,15.5,98.0,1.1,0.995,102.0,0.75,5.2,15.2

— WHY IS THIS NEEDED?
Bio-potency control is critical for peptide drugs (Novo Nordisk, Eli Lilly, Gan & Lee):
• GLP-1 agonists must effectively bind to receptor and activate cAMP cascade
• Post-translational modifications (deamidation, oxidation) may reduce activity without mass change
• Bioassay is "gold standard" for confirming functionality of complex molecule
• Required by regulators for release of each biological batch

⚠️ CRITICAL:
• Relative potency 80-125% — standard range for bioequivalence
• EC50 must be reproducible between runs (within 30% of standard)
• Emax ≥90% confirms drug is full agonist
• R² ≥0.98 ensures reliability of dose-response model
• Z-Factor ≥0.5 indicates high signal separation quality in plate
• CV ≤15% ensures statistical significance of results

Key features:
• Dose-response curve quality assessment (Hill equation fit)
• Relative potency control against reference standard
• Bioassay quality validation (Z-factor, CV)
• Support for various peptides (Semaglutide, Liraglutide, Tirzepatide)

Critical parameters:
• Relative potency: 80–125%
• EC50: Within 30% of standard
• Emax: ≥90%
• R²: ≥0.98
• Z-Factor: ≥0.5
• CV: ≤15%

💡 Usage tips:
1. Always include reference standard in each plate run for normalization
2. Use 4-parameter logistic model for curve fitting
3. High Z-factor (>0.7) is desirable for screening assays
4. Compare EC50 of new batches with historical data to detect trends
5. If low Emax observed, check for aggregates or drug degradation

⚠️ Note: Unlike chemical tests, bioassays have high variability. The utility helps distinguish real product quality issues from experimental artifacts (poor fit, high noise). For dual agonists (like Tirzepatide), separate analysis for each receptor (GLP-1R and GIPR) is required.

Traceability and limitations

• The URS is used as the source document for functional specification, CSV review and later validation work.
• This document does not replace an approved pharmacopoeial monograph, validated analytical method or internal product specification.
• For product-specific limits, the approved customer specification takes priority.

The functional specification describes the behaviour of the standalone FUZKK console utility, input-data format, evaluation algorithm and output JSON structure.

Functional flow

1. Main() checks the number of arguments.
2. If no arguments are provided: PrintHello() prints the description and built-in input.csv example, then RunDemoEvaluation() executes Evaluate() over GetDemoData() and prints demo JSON.
3. If two arguments are provided: RunWithFiles(input.csv, output.json) reads CSV, evaluates each record and writes LabWare-compatible JSON.
4. LoadData() uses CultureInfo.InvariantCulture and shall not be called in no-arguments mode.
5. Evaluate() returns a named tuple with BatchNumber, ProductName, Parameters, CriticalFailCount, WarningCount, Recommendation and RecommendationEN.
6. GetIssues() builds messages for ErrorMessage in WARNING/FAIL cases.

Evaluation rules

• PASS: CriticalFailCount = 0 and WarningCount = 0.
• WARNING: CriticalFailCount = 0 and WarningCount > 0.
• FAIL: CriticalFailCount > 0.
• ERROR: exception during reading or processing.
• ErrorMessage remains empty for PASS.
• Limits are embedded in Program.cs; no external limit configuration is required.

Input and fields

BatchNumber,ProductName,EC50_pM,Emax_Percent,HillSlope,R_Squared,RelativePotency_Percent,Z_Factor,CV_Percent,StandardEC50_pM
GLP1-BIO-2026-001,Semaglutide API,15.5,98.0,1.1,0.995,102.0,0.75,5.2,15.2
GLP1-BIO-2026-002,Liraglutide Bulk,250.0,95.0,1.0,0.990,98.0,0.70,6.5,245.0
GLP1-BIO-2026-003,Tirzepatide API,8.2,99.0,1.2,0.998,105.0,0.80,4.0,8.0

Output JSON

{
  "Header": {
    "UtilityName": "GLP1_Potency_Bioassay_Reviewer",
    "Version": "1.0.0",
    "Timestamp": "UTC",
    "InstrumentID": "FUZKK-QC-WORKSTATION",
    "OperatorID": "Admin"
  },
  "Samples": [
    {
      "SampleID": "from BatchNumber",
      "BatchNumber": "from CSV",
      "ProductName": "from CSV",
      "TestName": "utility-specific test",
      "AnalysisCode": "utility-specific code",
      "Status": "PASS | WARNING | FAIL | ERROR",
      "StatusCode": "1 | 2 | 0 | -1",
      "ErrorMessage": "",
      "Description": "Russian recommendation",
      "DescriptionEN": "English recommendation",
      "Results": [
        {
          "ParameterName": "parameter",
          "ResultValue": 0.0,
          "UnitOfMeasure": "unit",
          "SpecificationLimit": "limit",
          "IsWithinSpec": true
        }
      ]
    }
  ]
}