Dulaglutide — analytical characterisation
Chromatographic conditions, identity, related substances, presentation, reconstitution and in-use stability.
§6Analytical characterisation
§6.1Chromatographic conditions
- Column
- Size-exclusion chromatography for aggregates; protein A or cation-exchange for charge variants; peptide mapping after enzymatic digestion for sequence confirmation
- Mobile phase and gradient
- Method-dependent; reverse-phase intact analysis is not the primary tool for a 60 kDa glycoprotein
- Detection
- UV 280 nm (aromatic content dominates at this size); fluorescence and multi-angle light scattering as appropriate
- Retention
- Not applicable in the small-peptide sense
§6.2Identity by mass spectrometry
Intact mass by high-resolution MS after deglycosylation; glycan profiling by released-glycan analysis. Average intact mass is heterogeneous because of glycosylation.[3]
§6.3Related substances and degradation
Table 7. Related substances recorded for Dulaglutide, with the process or storage route that generates each and its analytical signature.
| Related substance | Origin | Analytical signature |
|---|---|---|
| Aggregates and high-molecular-weight species | Process or storage | Size-exclusion chromatography; a critical quality attribute for any Fc-fusion |
| Charge variants | Deamidation, C-terminal lysine heterogeneity | Ion-exchange chromatography |
| Clipped species | Proteolysis during production | Reduced capillary electrophoresis or peptide mapping |
| Host-cell protein and residual DNA | Upstream process | Immunoassay and quantitative PCR; not applicable to synthetic peptides |
Degradation routes
- Aggregation
- Deamidation and isomerisation
- Oxidation of methionine in the Fc region
- Fragmentation at the hinge
§7Presentation, reconstitution and storage
§7.1Presentation and reconstitution
- Presentation
- Aqueous solution in single-dose pen or prefilled syringe
- Reconstitution
- Not applicable
- Storage, lyophilised
- Not applicable
- Storage, reconstituted
- 2–8 °C in original carton, protected from light
- In-use period
- May be kept at up to 30 °C for a maximum of 14 days
Freezing destroys the product. Because the molecule is a glycoprotein, the analytical and stability framework is that of a biologic, and comparability rather than chemical equivalence is the relevant regulatory concept for any follow-on product.
§7.2In-use stability
Applicable standards: CEI-MS-01 · CEI-MS-02 · CEI-MS-03 · CEI-MS-04 · CEI-MS-05 · CEI-MS-06. The full series is at methodological standards.
Working calculators: reconstitution and insulin-unit conversion · purity against peptide content · certificate minimum-data checker.
References cited on this page
References are numbered in order of first citation in this document. Each superscript in the text links to its entry below.
- United States Pharmacopeial Convention. General Chapter ⟨1225⟩ Validation of Compendial Procedures. United States Pharmacopeia — National Formulary (USP–NF) 2024;USP 2024 Issue 1. identifier not held by the Institute
- International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use. ICH Q2(R2) Validation of Analytical Procedures. ICH Harmonised Guideline 2023;Step 4 version, 1 November 2023. identifier not held by the Institute
- International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use. ICH Q6B Specifications: Test Procedures and Acceptance Criteria for Biotechnological/Biological Products. ICH Harmonised Tripartite Guideline 1999;Step 4 version. identifier not held by the Institute
- International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use. ICH Q1A(R2) Stability Testing of New Drug Substances and Products. ICH Harmonised Tripartite Guideline 2003;Step 4 version. identifier not held by the Institute
- Manning MC, Chou DK, Murphy BM, Payne RW, Katayama DS. Stability of protein pharmaceuticals: an update. Pharmaceutical Research 2010;27(4):544–575. doi:10.1007/s11095-009-0045-6 · PMID 20143256
Identifiers are reproduced only where the Institute holds them. Where a digital object identifier or PubMed identifier is not shown, the Institute has recorded the journal and year and has not constructed an identifier.