MMS tracks stability and structure to assure the safety, efficacy and functionality of biotherapeutics

Meet Apollo, by RedShiftBio — your next-gen solution to the challenges of protein characterisation.

Apollo is RedShiftBio’s second-generation, fully automated flagship system designed to enable researchers to detect previously undetectable changes in biomolecule higher-order structure for analysis of formulations, stability, and lot-to-lot reproducibility. Measurement of these small changes allows the identification of the molecules, formulations, and conditions which are most likely to lead to stable drug products.

Apollo is powered by Microfluidic Modulation Spectroscopy (MMS) and provides ultra-sensitive, ultra-precise structural analysis of a wide range of biomolecules. When compared to CD or FTIR, MMS can detect structural change 20x faster and with 30x greater sensitivity*. MMS combines a high-power Quantum Cascade Laser with real-time buffer referencing. This provides the power to analyse both low- and high-concentration samples, in formulation buffer without excipient interference, and with the ability to detect small but critical structural changes. MMS delivers accurate and reproducible measurements from 0.1 mg/mL to >200 mg/mL under any conditions relevant to your study.

These unique features of MMS enable significant workflow improvements in different stages of biopharma development. In discovery, MMS can add more robust selection criteria for candidate screening by incorporating structural/stability monitoring earlier in the development process. In formulation, MMS has been shown to eliminate costly downstream failures by automatically analysing and comparing samples across a study to predictively identify optimal buffer formulations, stability profiles, and ideal storage conditions. Additionally, MMS tracks stability and structure — Critical Quality Attributes — across the entire manufacturing process, guaranteeing the safety, efficacy, and functionality of biotherapeutics.

Microfluidic Modulation Spectroscopy in Action

Researchers at Northeastern University recently demonstrated the utility of MMS to characterise IgG light chains found in patients with amyloidosis as part of an effort to elucidate the aggregation mechanism and provide therapeutic insights. In this study, MMS was employed in direct comparison to CD to study the aggregation propensity of samples based on three different sources of IgGs (Light and variable chain IgGs from amyloid-disease positive, disease-prone, and disease negative samples). MMS interrogates the Amide I band of the IR spectrum to sensitively probe protein structure while modulating against the reference buffer for accurate background subtraction in aqueous-based samples. Compared to CD, MMS can much more accurately measure beta-sheet structures and distinguish between native beta-sheets (intramolecular) and aggregated beta-sheets (intermolecular), a distinction that is critical for monitoring amyloid formation progression. General qualitative trends were observed in the CD structural results, but due to poor sensitivity for beta-sheet content, no quantitative conclusions could be made.

MMS provided quantitative HOS characterisation and the ability to distinguish between native and aggregation-prone beta-sheet structures, which are critical for amyloidogenic protein studies.

MMS proved its value to formulation workflows in another recent example by directly measuring buffer-induced changes in protein structure and folding — a rapidly growing area of interest due to structure being a key quality attribute in biopharmaceutical development. In this instance, MMS characterised buffer-induced structural differences of lysozyme, a well-characterised alpha-helix-rich protein, in water and three common buffers: Phosphate Buffer (PB), Phosphate Buffered Saline (PBS), and Tris buffer. Absorption spectra in the Amide I region were automatically collected and processed to calculate higher order structure (HOS) percentages and the overall structural similarities between samples in all prepared conditions. The results showed the enzyme exhibited various degrees of structural change within these buffers, and that these changes could be quantified to inform buffer selection decisions to support ideal protein activity.

To request a demo, proof-of-concept, or a discussion on how MMS can enhance your research or product development goals, please contact ATA Scientific at enquiries@atascientific.com.au or +61 2 9541 3500.

*Journal of Pharmaceutical Sciences, JAN 2020