Analytical Chromatography for Protein Analysis

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Advanced analytical chromatography solutions for protein analysis

Biotherapy manufacturers, especially those producing monoclonal antibodies (mAbs), rely heavily on chromatography for its precision and accuracy. Chromatography is an ideal technology to leverage in maintaining the purity, potency, and safety of these vital therapeutic agents. Techniques like High-Performance Liquid Chromatography (HPLC), Ion-Exchange Chromatography (IEX), and Size-Exclusion Chromatography (SEC) are crucial for detecting impurities, assessing molecular characteristics, and quantifying active ingredients.

Chromatography also helps identify protein aggregates and provides detailed insights into the molecular structure and charge variants of mAbs. This thorough analysis maintains consistency and reproducibility across batches, enhancing product quality and supporting regulatory compliance.

Moreover, chromatography streamlines the manufacturing process. It allows manufacturers to monitor production stages, identify issues early, and make necessary adjustments, thereby improving yield and product quality. Integrating chromatography into analytical applications is essential for manufacturers to deliver high-quality therapeutic agents to the market.

Analytical testing in the bioprocessing workflow

Analytical testing is crucial in the bioprocessing workflow to support the quality and safety of biopharmaceuticals. It helps detect impurities, assess molecular characteristics, and quantify active ingredients, maintaining consistency and reproducibility across batches. This rigorous testing supports regulatory compliance and contributes to overall product quality.

Thermo Fisher Scientific offers a robust portfolio of testing solutions to help simplify analytical processes while helping you meet or exceed regulatory guidelines.

Analytical chromatography for protein characterization

Throughout the manufacturing process, analytical chromatography is employed to assess the molecular characteristics, quantify active ingredients, and support consistency and reproducibility across product batches.

  • Monitors cell line productivity
  • Enables effective scalability
  • Supports downstream processing efficiency
  • Reduces therapeutic impact
  • Measures concentration in cell supernatant
  • Helps determine binding capacity
  • Enables process optimization
  • Supports process monitoring
  • Measures antibody titer in final formulation

Selecting an HPLC column

When choosing the right HPLC column for your analytical application, consider several key factors. The molecular size and characteristics must be considered when selecting pore size and surface chemistry to help ensure optimal separation. The material and stability of the column are also crucial, as they need to withstand the mobile phase conditions and remain reliable through repeated use. Additionally, the column's resolution and efficiency are important for achieving accurate and reproducible results. Finally, regulatory compliance and validation requirements should guide the selection process to help ensure that the column meets the necessary standards for use in mAb production and quality control.

Protein analytics HPLC solutions

mAb affinity columns

Discover our specialized HPLC columns designed for the high-resolution separation and analysis of monoclonal antibodies based on their affinity interactions.

Protein affinity columns

Find HPLC columns designed to separate and analyze proteins based on their specific binding interactions with ligands immobilized on the column matrix.

General purpose HPLC columns

Explore general purpose analytical columns, including ion exchange, reverse phase, and activated types, for versatile separation and analysis of diverse chemical compounds.

Discover the power of POROS resins

Thermo Scientific POROS resins are engineered to deliver fast, high-resolution analytical separations, supporting accurate and reproducible characterization of biomolecules. Built on a robust polymeric matrix, POROS technology offers exceptional mechanical stability with high flow rates, with consistent performance even under demanding HPLC conditions.

The unique bead structure features a large surface area that enables rapid mass transfer, facilitating reliable charge variant analysis, aggregate detection, and molecular profiling—all with greater speed and efficiency. With proven versatility across analytical applications from development through manufacturing, POROS chemistries empower analysts to generate actionable data quickly and with confidence, supporting quality and consistency at every stage.

Attributes of POROS chromatography resins:

  • 50-micron particle size—High-resolution separations with sharp peak shapes and reduced elution volumes, facilitating precise molecular characterization in complex analytical workflows
  • Large interconnected throughpores—Help minimize mass transfer resistance, enabling rapid separation while maintaining capacity and resolution across a broad range of flow rates
  • Rigid polystyrene-divinylbenzene backbone—Predictable, scalable performance with outstanding robustness and chemical stability, supporting consistent results under demanding analytical conditions

Help elevate analytical confidence with CaptureSelect Ligands

Thermo Scientific CaptureSelect affinity ligands offer exceptional specificity and selectivity to analytical workflows, enabling targeted detection and quantification of diverse biologics—from full-length antibodies to antibody fragments and next-generation therapeutics. Built on innovative single-domain antibody technology, CaptureSelect chemistries facilitate precise binding, reproducibility, and efficiency, simplifying complex assays while improving accuracy and reliability.

Well suited for challenging biological samples, CaptureSelect ligands help enhance analytical column performance by facilitating clean, consistent separations and dependable results. When paired with POROS bead technology, they enable highly specific affinity separations and molecular characterization that support the quality, consistency, and scalability required from development through manufacturing.

Benefits of CaptureSelect ligands:

  • Camelid-derived single-domain affinity ligands—Compact (~12–15 kDa) VHH ligands produced in an animal-origin–free yeast system for high specificity, affinity, and stability, supporting accurate HPLC analysis of complex biologics
  • Mild elution conditions—CaptureSelect ligands enable gentle release of target molecules during HPLC workflows, designed to protect protein integrity and facilitate reliable characterization
  • Consistent performance and supply—Manufactured under ISO-certified conditions with excellent lot-to-lot reproducibility, CaptureSelect ligands offer dependable results for routine and high-throughput HPLC analyses

Request a technical consultation

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Analytics Knowledge Hub


Explore the Analytics Knowledge Hub and discover a robust collection of webinars, articles, eBooks, infographics, and other digital resources to help you optimize analytical testing in your bioprocess workflow.
 

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