A Review of Emerging Analytical Technologies for Biologics: From Microfluidic Modulation Spectroscopy to Next-Generation Structural Characterization

Biologics, including monoclonal antibodies, recombinant proteins, and antibody–drug conjugates, form a rapidly expanding class of therapeutics whose safety and efficacy rely on maintaining precise higher-order structures (HOS). Conventional analytical techniques such as circular dichroism, Fourier-transform infrared spectroscopy, and differential scanning calorimetry provide valuable but often limited insight into protein conformation. Recent advances in microfluidic modulation spectroscopy (MMS) have enabled high-resolution structural characterization with minimal sample consumption under near-native conditions. MMS offers rapid, sensitive detection of secondary structure changes and is increasingly applied in formulation screening, stability testing, and comparability assessments. Complementary next-generation tools—such as cryo-electron microscopy, hydrogen–deuterium exchange mass spectrometry, native mass spectrometry, and mass photometry—further enhance the analytical depth by revealing conformational dynamics and molecular interactions at multiple structural levels. The integration of these experimental techniques with computational approaches like AlphaFold and Rosetta provides predictive models for protein folding and stability. Together, these technologies form a powerful multimodal analytical framework that supports the principles of Quality by Design (QbD) and regulatory expectations for biosimilarity and consistency in biopharmaceutical development. By combining precision spectroscopy, high-throughput workflows, and computational modeling, MMS and allied techniques are transforming structural characterization, enabling faster and more reliable development and quality assurance of biologics worldwide.

کلیدواژه ها:

Biologics ، Microfluidic Modulation Spectroscopy (MMS) ، Higher-Order Structure (HOS) ، Critical Quality Attributes (CQAs) ، Cryo-Electron Microscopy ، HDX-MS