Leaders in characterization

Significance of the topic

The development and manufacturing of therapeutic proteins require precise characterization to ensure product integrity and efficacy. Advanced analytical methods such as high-resolution mass spectrometry (HRMS) combined with liquid chromatography (LC) enable detailed profiling of protein forms, detection of variants, and control of quality attributes, which are critical for drug development and diagnostic applications.

Objectives and study overview

This case study outlines how R&D Systems, a Bio-Techne brand, integrated state-of-the-art LC-MS technology into its protein development workflows under cGMP conditions. The primary goal was to enhance in-house capabilities for comprehensive structural analysis, accelerate problem solving in manufacturing and storage issues, and deliver faster, high-confidence results to internal and external customers.

Methodology and Instrumentation

R&D Systems deployed a Thermo Scientific platform tailored for protein analysis:

• Vanquish Flex UHPLC system for biocompatible, high-precision separations with Viper Fingertight Fittings
• Q Exactive HF Hybrid Quadrupole-Orbitrap mass spectrometer with BioPharma Option, offering High Mass Range (HMR), Protein, and Standard modes
• BioPharma Finder software facilitating intact mass analysis, top/middle-down sequencing, peptide mapping, and data deconvolution
• MAbPac SEC-1 size exclusion HPLC column for high-resolution separation of monoclonal antibodies and protein complexes

Key results and discussion

Implementation of the HRAM LC-MS workflow enabled same-day structural insights into complex proteins. In one example, size exclusion chromatography-mass spectrometry (SEC-MS) identified an oxidized monomer variant in a TNF-alpha trimer preparation, pinpointing storage conditions as the cause. The integrated platform delivered unambiguous mass measurements and accelerated troubleshooting, replacing previous low-resolution methods and outsourcing approaches.

Benefits and practical applications of the method

• Enhanced confidence in mass accuracy and variant identification at both peptide and intact protein levels
• Reduced turnaround times with in-house analysis replacing outsourced services
• Streamlined workflows supporting rapid decision making in process development and quality control
• Scalable solution adaptable to a broad molecular weight range from small cytokines to full-length antibodies

Future trends and potential applications

As biologics continue to evolve, multi-attribute methods combining HRAM MS and advanced chromatography are expected to become standard for real-time monitoring of critical quality attributes. Integration of automated data processing, increased throughput, and online process analytics will further accelerate protein characterization in research and manufacturing environments.

Conclusion

By investing in a comprehensive LC-MS platform and expert analytical teams, R&D Systems significantly improved the speed, precision, and scope of protein characterization. This approach ensures consistent product quality, supports rapid problem resolution, and strengthens the company’s position in the competitive biologics market.

Reference

• Bio-Techne Corporation. Case Study 73813: Delivering protein product quality through characterization expertise.