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Analysis of Protein Aggregates Concentration and Morphology by Sedimentation Equilibrium (SE-AUC) (CAT#: STEM-B-0297-CJ)

Introduction

The generic term 'aggregates' refers to species characterized by a wide size range, diverse morphologies and structures. Protein aggregates may start in the low nanometer size range but then can grow into the micrometer and even visible size range.<br /><br />Most protein therapeutics and many other biopharmaceutical compounds are inherently unstable and can undergo aggregation through various pathways. Aggregates of various kinds can be formed, such as reversible and non-reversible, soluble, and non-soluble etc. In addition,Aggregation maybe occur because of exposure to air-liquid or liquid-solid interfaces, e.g., during mixing, during filling and shipping, during reconstitution of lyophilized products, or through contact with chromatography columns, pumps, pipes, vessels, filters, etc. Aggregation can directly influence the efficacy of the therapy by reducing the number of functional molecules, but also indirectly influence efficacy as well as safety of a therapy by inducing side-effects, such as unwanted immunogenicity.<br /><br />Molecular weight, also called molecular mass, mass of a molecule of a substance, based on 12 as the atomic weight of carbon-14. It is calculated in practice by summing the atomic weights of the atoms making up the substance's molecular formula.




Principle

SE-AUC uses the same physical principles and detection mechanisms as SV-AUC, but with a crucial difference: it is performed at a lower rotation speed where back-diffusion – from the bottom of the cell (high concentration) towards the top (low concentration) – becomes significant. Over time, this back-diffusion reaches an equilibrium with the sedimentation movement and forms a stable concentration profile. For an extended period, the formation and stable position of this profile is recorded. This delivers most accurate information on the molecular weight of the species in solution because the molecule-shape is not influencing the results of SE-AUC as it is the case for HP-SEC, AF4 or SV-AUC.

Applications

Biopharmaceutica

Procedure

1. Sample preparation.
2. Place the sample and set up the experiment according to the Sedimentation Velocity Analytical Ultracentrifugation (SV-AUC) instrument instructions.
3. Analyse the data.

Materials

• Sample: peptides, proteins, viruses, nanomedicines and more.
• Equipment: Sedimentation Equilibrium (SE-AUC) Instrument

Notes

SE-AUC is currently the gold standard method for determining molecular weights of macromolecules in their native buffer solution.