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Analysis of Protein Aggregates Conformation by Analytical Ultra-Centrifugation (CAT#: STEM-B-0306-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.

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.

Molecular weight, also called molecular mass, mass of a molecule of a substance, based on 12 as the atomic weight of carbon-12. It is calculated in practice by summing the atomic weights of the atoms making up the substance's molecular formula.




Principle

Analytical ultracentrifugation – or more precisely the sedimentation velocity (SV-AUC) and sedimentation equilibrium (SE-AUC) method – is based on a simple principle: when (macro)molecules in solution are subject to a centrifugal force, they begin to settle at a certain velocity. With a mathematical correlation between sedimentation behavior and hydrodynamic properties, scientists can determine many critical parameters of a drug substance or drug product for a variety of biopharmaceutical applications from analyzing small peptides to investigating macromolecular interactions.

Applications

Biopharmaceutica

Procedure

The AUC concept consists in applying a centrifugal force, which allows sedimentation, and, therefore, the protein separation. The Proteins that aggregate faster are sedimented first, while remain in solution those proteins that aggregate slowly.

Materials

• Sample: Proteins
• Equipment: Analytical Ultra-Centrifugation

Notes

• The formation of aggregates in your biopharmaceutical product can have a negative effect on safety, efficacy and function. Regulatory authorities expect that orthogonal characterization techniques are used to fully understand the aggregation profile of any molecule.
• Measurable range: 1–100 nm
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