Workflow Solutions for Peptide Therapeutics - Application Compendium

Importance of the Topic

Peptide therapeutics have emerged as a versatile class of biologically active drugs, combining high specificity, favorable safety profiles, and low toxicity. Among these, glucagon-like peptide-1 (GLP-1) agonists play a key role in treating type 2 diabetes and obesity by modulating insulin and glucagon secretion. Rigorous analytical characterization throughout research, development, and manufacturing is critical to ensure product quality, efficacy, and regulatory compliance.

Objectives and Overview

This compendium presents an end-to-end suite of analytical workflows tailored for synthetic and recombinant peptide drugs, with a special focus on GLP-1 therapeutics. It reviews impurity profiling, purity assays, identity confirmation, structural analysis, aggregate detection, stability testing, and quantitative bioanalysis. The document also highlights Agilent’s integrated solutions designed to address USP, ICH, and pharmacopeial requirements.

Methodology and Instrumentation

Key analytical techniques include:

• Reversed-phase LC (RP-LC-UV, RP-LC/MS) for purity and impurity testing
• LC/MS and LC/MS/MS for mass determination and sequence confirmation
• Size-exclusion chromatography (SEC) and 2D-LC for aggregate profiling
• Ion mobility–MS (IM/MS) for conformational studies
• GC/MS and headspace GC for residual solvents
• ICP-MS for elemental impurities
• Raman and FTIR spectroscopy for raw-material identity verification
• Auto-derivatization LC for amino acid composition

Representative instrumentation comprises Agilent 1290 Infinity II/III LC systems, AdvanceBio columns, 6545XT Q-TOF, 6495D Triple Quadrupole, 1260 Prime Bio LC, 8890 GC/MS, 7800 ICP-MS, Vaya Raman, and SLIMS laboratory informatics.

Main Results and Discussion

Agilent workflows demonstrated robust scale-up from analytical to preparative peptide purification, precise mapping of post-translational modifications via electron capture dissociation, and aggregate identification under MS-compatible conditions. Stability studies identified key degradation products, while MRM-based assays enabled quantitation of native and oxidized tirzepatide down to 0.025 ng/mL. Automated derivatization and data deconvolution streamlined amino acid composition and intact mass confirmation.

Benefits and Practical Applications

• Comprehensive monitoring of critical quality attributes (CQAs)
• Regulatory compliance with USP <1503>, Ph. Eur., and ICH guidelines
• High sensitivity and reproducibility for impurity profiling
• Seamless method transfer and automation for high throughput
• Inorganic and organic impurity detection in a single platform

Future Trends and Opportunities

Analytical demands will grow with generic peptide approvals and complex modalities. Anticipated trends include deeper integration of artificial intelligence for method development, expanded use of high-resolution MS and 2D-LC, advanced ion mobility for structural insights, and fully automated, end-to-end digital workflows for real-time quality control.

Conclusion

Agilent’s integrated portfolio—from sample prep and chromatography to MS detection and informatics—provides a streamlined, compliant framework for peptide therapeutic analysis. By combining high-performance instruments, specialized columns, and software-driven automation, laboratories can achieve reliable, high-throughput characterization at every development stage.

Instrumentation

• LC Systems: 1290 Infinity II/III, 1260 Infinity Prime Bio, 1290 bio 2D-LC
• MS Platforms: 6545XT AdvanceBio Q-TOF, 6495D Triple Quadrupole, InfinityLab LC/MSD iQ/XT
• GC/MS: 8890 GC, 5977B MSD, 7697A Headspace Sampler
• ICP-MS: 7800
• Spectroscopy: Vaya Raman, Cary FTIR, RapID system
• Automation: AssayMAP Bravo, SLIMS LIMS/ELN

References

• Figure 4 | Biological Activities of GLP-1. Omics Online.