CE_Glycosylation_Profiler

URS & FS — User Requirements and Functional Specification

This document describes the controlled interface, user requirements and functional behaviour of CE_Glycosylation_Profiler. The utility is intended for automated verification of laboratory, pharmacopoeial, analytical or manufacturing QC parameters using input.csv and producing a structured output.json result.

Domain limits and critical parameters

• • Asialo forms: ≤5.0%
• • Isoform distribution (G0–G4): within specification
• • Sialic Acid Index: ≥2.5
• • Sialic acids at the ends of glycan chains protect the protein from degradation and removal from the bloodstream.
• • Capillary Electrophoresis (CE) with Laser-Induced Fluorescence (LIF) allows separation of isoforms by charge (number of sialic acids).
• • Glycosylation profile affects potency, pharmacokinetics, and immunogenicity.
• • Control is necessary to ensure consistency of the manufacturing process (cell culture consistency).
• • Asialo forms (G0) must be minimal. Their accumulation indicates purification issues or protein instability.
• • Sialic Acid Index (SA Index) is an integral quality indicator. Drop below 2.5 is critical for EPO.
• • Shift of profile towards less sialylated forms (left shift on electropherogram) is a sign of degradation or cell-line glycosylation errors.
• • Method requires high resolution and use of standards for migration time calibration.
• • Calculation of glycoform percentage distribution.
• • Assessment of Sialic Acid Index.
• • Control of recombinant protein expression process quality.
• • Asialo Forms: ≤5.0%
• • Disialo/Tetrasialo Forms: Within Specification Range

URS — User Requirements Specification

input.csv contract

BatchNumber,Asialo%,Mono%,Di%,Tri%,Tetra%,SA_Index
EPO-CE-2026-001,1.5,15.0,45.0,25.0,13.5,2.8
EPO-CE-2026-002,2.0,18.0,42.0,24.0,14.0,2.7
EPO-FAIL-DESIALO,8.5,25.0,40.0,15.0,11.5,2.1

FS — Functional Specification

Example output.json

{
  "utilityId": "ce-glycosylation-profiler",
  "utilityName": "CE_Glycosylation_Profiler",
  "overallStatus": "PASS|WARNING|FAIL",
  "sourceFile": "input.csv",
  "checks": [
    {
      "parameter": "BatchNumber",
      "value": "EPO-CE-2026-001",
      "status": "PASS|WARNING|FAIL",
      "message": "Rule-based check result"
    },
    {
      "parameter": "Asialo%",
      "value": "1.5",
      "status": "PASS|WARNING|FAIL",
      "message": "Rule-based check result"
    },
    {
      "parameter": "Mono%",
      "value": "15.0",
      "status": "PASS|WARNING|FAIL",
      "message": "Rule-based check result"
    },
    {
      "parameter": "Di%",
      "value": "45.0",
      "status": "PASS|WARNING|FAIL",
      "message": "Rule-based check result"
    },
    {
      "parameter": "Tri%",
      "value": "25.0",
      "status": "PASS|WARNING|FAIL",
      "message": "Rule-based check result"
    },
    {
      "parameter": "Tetra%",
      "value": "13.5",
      "status": "PASS|WARNING|FAIL",
      "message": "Rule-based check result"
    },
    {
      "parameter": "SA_Index",
      "value": "2.8",
      "status": "PASS|WARNING|FAIL",
      "message": "Rule-based check result"
    }
  ],
  "criticalFindings": [],
  "warnings": [],
  "generatedFor": "QA/QC review and LIMS integration"
}

Traceability matrix

OQ/PQ test scenarios

QA/QC and change control

• Do not rename columns without updating the validator, documentation and test set.
• Retain input.csv, output.json, executable version, documentation and checksum.
• Before production use, perform IQ/OQ/PQ or equivalent CSV/CSA verification.
• Critical limits must be verified against the approved specification, local SOPs and registration dossier.