Literature

Orthogonal method in pharmaceutical analysis

by Thomas Kofoed, CEO, and Jette Friis Thirup, Head of Business Development, Alphalyse

Implementing at least one orthogonal method is essential for ensuring the precision of analytics in the pharmaceutical and biopharmaceutical industries. The approach increases the reliability of analytical techniques in ensuring product quality, safety, and regulatory compliance.

What is an orthogonal method?

The orthogonal method approach uses multiple independent techniques to analyze or validate results. By eliminating the reliance on a single method, this approach reduces potential biases and enhances the overall accuracy of measurements. In pharmaceuticals, orthogonality is essential for characterizing complex biological products, such as monoclonal antibodies (mAbs), vaccines, and cell or gene therapies.

Orthogonality in pharmaceutical quality control

The complexity of modern biologics necessitates robust analytical strategies. For instance, products incorporating nanomaterials or advanced delivery systems present unique challenges in ensuring consistent quality and safety. Since traditional analytical methods are often unsuited for these products, regulatory agencies like the FDA and EMA recommend orthogonal analytical techniques - such as mass spectrometry - to address the challenges effectively.

Measuring Critical Quality Attributes (CQAs)

Critical Quality Attributes (CQAs), such as protein aggregation levels or residual impurities, are central to product efficacy and safety. An orthogonal method allows pharmaceutical developers to measure CQAs with greater confidence. For example, combining ELISA (Enzyme-Linked Immunosorbent Assay) with Liquid Chromatography-Mass Spectrometry (LC-MS) provides a multi-faceted understanding of protein impurities, ensuring no critical components are overlooked.

Addressing regulatory demands

Orthogonal techniques are vital in meeting the tight requirements of regulatory bodies. For example, mass spectrometry (MS) is increasingly recognized for its ability to quantify Host Cell Proteins (HCPs) and other process-related impurities. When paired with complementary methods like ELISA, mass spectrometry improves sensitivity and provides detailed insights into individual impurities - a necessity for regulatory documentation and product approval.

 

ELISA-MS is a coverage method based on mass spectrometry

Technological contributions of MS as an orthogonal method

Advanced Mass Spectrometry

Mass spectrometry, particularly Data Independent Acquisition (DIA) methods such as SWATH-MS, exemplifies the power of orthogonality in analytical sciences. These techniques enable comprehensive identification and quantification of trace impurities. Unlike conventional methods, which may miss certain protein contaminants, orthogonal approaches ensure complete impurity profiling.

Supporting biopharmaceutical innovation

Biopharmaceuticals, such as cell and gene therapies, often involve complex manufacturing processes. As an orthogonal method, mass spectrometry plays a pivotal role in these processes, enabling detailed analysis at every stage - from early development to final product validation. Implementing mass spectrometry can provide the knowledge necessary to ensure that therapeutic products meet the highest safety and efficacy standards.

Final thoughts

The Orthogonal Method approach is more than just technical; it is a fundamental strategy for ensuring the reliability and robustness of pharmaceutical and biopharmaceutical products. By leveraging the independence and complementarity of analytical techniques, this approach reduces biases, enhances precision, and meets stringent regulatory expectations.

With increasingly complex therapeutics under development, adopting orthogonal strategies is not just advantageous, but essential. Whether ensuring advanced biologics' safety or meeting global health crises' demands, orthogonality underpins innovation, compliance, and trust in pharmaceutical development.

However, even for mAb products and recombinant products, orthogonal HCP analysis techniques can be critical to implement, as ELISAs may miss important protein impurities.

FAQ

Implementing an orthogonal method means using two or more independent analytical techniques to validate results. This approach is critical for ensuring reliability and precision in complex analyses, particularly in industries like pharmaceuticals, where accurate measurement of impurities and product consistency is essential.

In pharmaceutical quality control, using an orthogonal method allows for more accurate measurement of critical quality attributes (CQAs). For instance, combining ELISA (Enzyme-Linked Immunosorbent Assay) with mass spectrometry provides a comprehensive view of impurities, reducing the risk of overlooking potentially harmful contaminants.

Orthogonal techniques commonly used include:

  • ELISA and Liquid Chromatography-Mass Spectrometry (LC-MS) for impurity profiling.
  • Analytical ultracentrifugation alongside size-exclusion chromatography for studying protein aggregates.
  • Complementary spectrometric and chromatographic methods for comprehensive impurity analysis.

Regulatory agencies like the FDA and EMA recommend orthogonal methods to ensure the robustness and accuracy of analytical results. These methods help meet stringent safety and efficacy standards by providing comprehensive, unbiased insights into product quality, especially for advanced therapeutics like monoclonal antibodies and gene therapies.

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