AssayTransferProtocolChecker

Assay Transfer Protocol

Liquid Biopsy жидкостная биопсия cfDNA ctDNA CTC exosomes NGS qPCR

Utility description: Assay Transfer Protocol

Assay Transfer Protocol Checker — Analytical Method Transfer Verification

ℹ️ Utility performs statistical assessment of analytical method transfer success between laboratories according to USP <1224> and ICH Q2(R1):
• Accuracy Comparison: t-test to detect systematic bias between sending and receiving units.
• Precision Comparison: F-test to check equality of variances (reproducibility).
• Absolute Difference: Check of percentage difference of means against established limit.
• Decision Making: Classification of result as Successful, Conditional, or Failed.

⚠️ IMPORTANT:
• Method transfer requires receiving laboratory to prove competence in executing the procedure.
• Statistical tests are sensitive to sample size (N).

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

Input format:
ProtocolID,AnalyteName,SendingUnit_Mean,SendingUnit_SD,SendingUnit_N,ReceivingUnit_Mean,ReceivingUnit_SD,ReceivingUnit_N,Max_Allowed_Diff_Percent

Example:
MT-001,Assay,99.5,0.8,6,99.2,0.9,6,2.0

📍 Scope of Application (Usage Where):
• R&D to QC Transfer: Validation of routine control.
• Inter-laboratory Comparisons: When moving production to another site.
• Outsourcing: Transfer of methods to contract laboratories (CRO).
• Regulatory Dossier: Justification of data comparability.

— WHY IS THIS NEEDED?
Guarantee that analysis results obtained at new site are comparable to original ones.
This is critically important for supply continuity and compliance with licensing requirements.
Automation of statistical calculations eliminates errors in manual t- and F-criterion calculation.

⚠️ CRITICAL:
• Sampling: Minimum number of replicates (usually N≥6) for statistical reliability.
• Variance Homogeneity: If variances differ significantly, standard t-test is inapplicable.
• Limits: Percentage difference must be defined before experiment start.
• Outliers: Outliers must be investigated before statistical analysis.

Key features:
• Automatic t-test and F-test calculation
• Percentage mean difference assessment
• Flexible confidence interval adjustment
• Report generation for method transfer protocol
• Compliance with USP <1224> requirements

Critical parameters:
• Mean Diff %: < Spec Limit
• F-Ratio: < Critical Value
• T-Test: No significant difference (p > 0.05)
• N: Sufficient sample size

💡 Usage tips:
1. Planning: Define acceptance criteria in transfer protocol before testing.
2. Training: Ensure receiving lab analysts are trained on the method.
3. Equipment: Check instrument calibration in both laboratories.
4. Documentation: Save raw data of all replicates.
5. Investigation: Conduct root cause analysis upon "Failed" status.

⚠️ Note: This utility is a statistical support tool. Final decision on method approval is made by Quality Unit Head based on full report.

input.csv

ProtocolID,AnalyteName,SendingUnit_Mean,SendingUnit_SD,SendingUnit_N,ReceivingUnit_Mean,ReceivingUnit_SD,ReceivingUnit_N,Max_Allowed_Diff_Percent
MT-2026-API-001,Assay of Drug X,99.5,0.8,6,99.2,0.9,6,2.0
MT-2026-IMP-002,Impurity A,0.15,0.02,6,0.25,0.05,6,10.0
MT-2026-DISS-003,Dissolution Q,85.0,1.5,6,84.5,1.6,6,5.0

URS & FS — User Requirements and Functional Specification

This document describes the controlled interface and behaviour of AssayTransferProtocolChecker for Assay Transfer Protocol Checker.

Domain limits URS input.csv FS IQ/OQ/PQ

Domain limits and critical parameters

Key fragments from the source description are shown below. Before production use, limits must be verified against the approved specification, registration dossier and local SOPs.

ℹ️ Utility performs statistical assessment of analytical method transfer success between laboratories according to USP <1224> and ICH Q2(R1):
• Accuracy Comparison: t-test to detect systematic bias between sending and receiving units.
• Precision Comparison: F-test to check equality of variances (reproducibility).
• Absolute Difference: Check of percentage difference of means against established limit.
• Inter-laboratory Comparisons: When moving production to another site.
This is critically important for supply continuity and compliance with licensing requirements.
⚠️ CRITICAL:
• Sampling: Minimum number of replicates (usually N≥6) for statistical reliability.
• Limits: Percentage difference must be defined before experiment start.
• Outliers: Outliers must be investigated before statistical analysis.
• Compliance with USP <1224> requirements
Critical parameters:
• Mean Diff %: < Spec Limit
• F-Ratio: < Critical Value
• T-Test: No significant difference (p > 0.05)

URS — User Requirements Specification

ID	Requirement	Criticality	Acceptance criterion
URS-001	The utility shall accept an input.csv file for Assay Transfer Protocol Checker with headers defined in the data contract.	High	The file is processed without manual header editing.
URS-002	The utility shall perform deterministic QC evaluation without machine learning and without probabilistic conformance decisions.	High	Identical input data, rule version and configuration produce reproducible results.
URS-003	The utility shall validate mandatory fields, data types, ranges, units and domain plausibility.	High	Schema, conversion and range errors are explicitly reported.
URS-004	The utility shall apply domain limits and rules from the description, approved specification, registration dossier and local SOPs.	High	Each check has PASS/WARNING/FAIL and a clear message.
URS-005	The utility shall generate output.json with machine-readable results, source values, warnings, failures and critical findings.	High	JSON is suitable for LIMS/ELN/MES integration and QA/QC review.
URS-006	The utility shall preserve traceability between batch/sample, input file, applied rules and final status.	High	Output contains identifiers, checked parameters and audit metadata.
URS-007	The documentation shall support IQ/OQ/PQ, CSV/CSA and review by internal QA or inspectors.	Medium	URS, FS, input/output contract and test scenarios are supplied with the utility.
URS-008	The utility shall be used as a QC decision-support tool and not as a substitute for approved specifications and QA/QP release decision.	Medium	Documentation states change control and limit-verification expectations.

input.csv contract

#	Field	Type	Sample	Purpose
1	`ProtocolID`	string / controlled vocabulary	`MT-2026-API-001`	Controlled input parameter for deterministic QC rules.
2	`AnalyteName`	string / controlled vocabulary	`Assay of Drug X`	Controlled input parameter for deterministic QC rules.
3	`SendingUnit_Mean`	integer / decimal	`99.5`	Controlled input parameter for deterministic QC rules.
4	`SendingUnit_SD`	decimal	`0.8`	Controlled input parameter for deterministic QC rules.
5	`SendingUnit_N`	integer / decimal	`6`	Controlled input parameter for deterministic QC rules.
6	`ReceivingUnit_Mean`	integer / decimal	`99.2`	Controlled input parameter for deterministic QC rules.
7	`ReceivingUnit_SD`	decimal	`0.9`	Controlled input parameter for deterministic QC rules.
8	`ReceivingUnit_N`	integer / decimal	`6`	Controlled input parameter for deterministic QC rules.
9	`Max_Allowed_Diff_Percent`	decimal	`2.0`	Controlled input parameter for deterministic QC rules.

ProtocolID,AnalyteName,SendingUnit_Mean,SendingUnit_SD,SendingUnit_N,ReceivingUnit_Mean,ReceivingUnit_SD,ReceivingUnit_N,Max_Allowed_Diff_Percent
MT-2026-API-001,Assay of Drug X,99.5,0.8,6,99.2,0.9,6,2.0
MT-2026-IMP-002,Impurity A,0.15,0.02,6,0.25,0.05,6,10.0
MT-2026-DISS-003,Dissolution Q,85.0,1.5,6,84.5,1.6,6,5.0

Input validation rules

ID	Field	Rule	Criticality
VR-001	`ProtocolID`	The field shall match an approved dictionary or accepted string representation.	High
VR-002	`AnalyteName`	The field shall match an approved dictionary or accepted string representation.	High
VR-003	`SendingUnit_Mean`	The field shall match an approved dictionary or accepted string representation.	High
VR-004	`SendingUnit_SD`	The field shall match an approved dictionary or accepted string representation.	Medium
VR-005	`SendingUnit_N`	The field shall match an approved dictionary or accepted string representation.	Medium
VR-006	`ReceivingUnit_Mean`	The field shall match an approved dictionary or accepted string representation.	Medium
VR-007	`ReceivingUnit_SD`	The field shall match an approved dictionary or accepted string representation.	Medium
VR-008	`ReceivingUnit_N`	The field shall match an approved dictionary or accepted string representation.	Medium
VR-009	`Max_Allowed_Diff_Percent`	The field shall match an approved dictionary or accepted string representation.	Medium

FS — Functional Specification

ID	Function	Implementation
FS-001	CLI execution	Support execution modes: demo mode without arguments and production mode input.csv output.json.
FS-002	CSV import	Read input.csv in UTF-8/CSV-compatible format and validate header and expected columns.
FS-003	Schema validation	Check mandatory fields, column count, unknown key fields and empty mandatory values.
FS-004	Type conversion	Convert numeric, flag and text values; invalid format is recorded as a row-level error.
FS-005	Domain rule engine	Apply rules for Assay Transfer Protocol Checker, including critical limits from the description and approved specification.
FS-006	Status aggregation	Produce final status: FAIL for critical failure, WARNING for non-critical deviation, PASS for conformance.
FS-007	JSON export	Write output.json with detailed checks, source values, warnings, failures and critical findings.
FS-008	Audit support	Keep result structure suitable for review, deviation investigation and calculation reproduction.
FS-009	Integration contract	Support the scenario LIMS/ELN/MES → input.csv → utility → output.json → portal/admin review.
FS-010	Error handling	Return explicit messages for missing file, empty CSV, invalid schema, output write failure and invalid format.

Example output.json

{
  "utilityId": "assaytransferprotocolchecker",
  "utilityFolder": "AssayTransferProtocolChecker",
  "package": "LiquidBiopsy",
  "overallStatus": "PASS|WARNING|FAIL",
  "sourceFile": "input.csv",
  "processedAtUtc": "2026-06-10T00:00:00Z",
  "checks": [
    {
      "parameter": "ProtocolID",
      "value": "MT-2026-API-001",
      "status": "PASS|WARNING|FAIL",
      "message": "Deterministic rule-based check result",
      "ruleReference": "FS-RULE-001"
    },
    {
      "parameter": "AnalyteName",
      "value": "Assay of Drug X",
      "status": "PASS|WARNING|FAIL",
      "message": "Deterministic rule-based check result",
      "ruleReference": "FS-RULE-002"
    },
    {
      "parameter": "SendingUnit_Mean",
      "value": "99.5",
      "status": "PASS|WARNING|FAIL",
      "message": "Deterministic rule-based check result",
      "ruleReference": "FS-RULE-003"
    },
    {
      "parameter": "SendingUnit_SD",
      "value": "0.8",
      "status": "PASS|WARNING|FAIL",
      "message": "Deterministic rule-based check result",
      "ruleReference": "FS-RULE-004"
    },
    {
      "parameter": "SendingUnit_N",
      "value": "6",
      "status": "PASS|WARNING|FAIL",
      "message": "Deterministic rule-based check result",
      "ruleReference": "FS-RULE-005"
    },
    {
      "parameter": "ReceivingUnit_Mean",
      "value": "99.2",
      "status": "PASS|WARNING|FAIL",
      "message": "Deterministic rule-based check result",
      "ruleReference": "FS-RULE-006"
    },
    {
      "parameter": "ReceivingUnit_SD",
      "value": "0.9",
      "status": "PASS|WARNING|FAIL",
      "message": "Deterministic rule-based check result",
      "ruleReference": "FS-RULE-007"
    },
    {
      "parameter": "ReceivingUnit_N",
      "value": "6",
      "status": "PASS|WARNING|FAIL",
      "message": "Deterministic rule-based check result",
      "ruleReference": "FS-RULE-008"
    },
    {
      "parameter": "Max_Allowed_Diff_Percent",
      "value": "2.0",
      "status": "PASS|WARNING|FAIL",
      "message": "Deterministic rule-based check result",
      "ruleReference": "FS-RULE-009"
    }
  ],
  "criticalFindings": [],
  "warnings": [],
  "audit": {
    "inputHash": "sha256:<calculated at runtime>",
    "rulesVersion": "<utility executable version>",
    "documentation": "AssayTransferProtocolChecker.documentation.html"
  }
}

Traceability matrix

URS	FS	Test	Evidence
URS-001	FS-001, FS-002	OQ-001	Verify execution and import of valid input.csv.
URS-002	FS-005, FS-006	OQ-004	Repeat the same dataset and compare output.json.
URS-003	FS-003, FS-004, FS-010	OQ-002, OQ-003	Verify missing columns and invalid types.
URS-004	FS-005, FS-006	OQ-004, PQ-001	Verify critical deviations on real/boundary data.
URS-005	FS-007, FS-009	OQ-005	Verify JSON schema and downstream-system suitability.
URS-006	FS-008	OQ-006	Verify identifiers and audit metadata.
URS-007	FS-008, FS-010	IQ-001, OQ-007	Verify documentation completeness and control evidence.
URS-008	FS-005, FS-008	PQ-002	Verify review workflow and no replacement of QA decision.

IQ/OQ/PQ test scenarios

ID	Scenario	Expected result
IQ-001	Verify executable, input.csv, documentation and checksum availability.	Delivery set is complete; version is recorded.
OQ-001	Valid sample row from input.csv.	PASS or acceptable WARNING according to rules.
OQ-002	Remove a mandatory CSV column.	Schema error or FAIL with missing-column reference.
OQ-003	Place a non-numeric value into a numeric field.	Type-conversion error with row/field reference.
OQ-004	Set a critical parameter outside the limit.	FAIL and critical finding.
OQ-005	Verify output.json structure.	All mandatory sections are present and JSON is valid.
OQ-006	Verify batch/sample traceability.	Input and result identifiers match.
PQ-001	Verify 3–5 real user batches/samples.	Result is confirmed by QC/QA review.
PQ-002	Verify deviation workflow and manual QA decision.	Utility supports review but does not replace approved decision.

QA/QC and change control

Do not rename columns without updating validator, documentation and test set.
Retain input.csv, output.json, executable version and checksum.
Before production use, perform IQ/OQ/PQ or equivalent CSV/CSA verification.
Critical limits shall be verified against the approved specification, registration dossier and local SOPs.
The utility provides structured QC decision support; final release decision remains with QA/QP and approved procedures.

Included in packages

Biopharmaceuticals Extended QC Suite

Extended QC utility coverage for mAbs, biosimilars, proteins, insulins, vaccines, mRNA/LNP, HCP, sterile release, microbiology, water, cleanroom and stability workflows.

Open

Liquid Biopsy QC Suite

QC and pre-analytical control package for liquid biopsy workflows: cfDNA/ctDNA, CTC, EV/exosomes, methylation, NGS/qPCR/ddPCR, sample quality, contamination, sensitivity and reporting checks.

Open