Role of analytical ultracentrifugation in assessing the aggregation of protein biopharmaceuticals
Journal Title: The AAPS Journal - Year 2006, Vol 8, Issue 3
Abstract
In developing and manufacturing protein biopharmaceuticals, aggregation is a parameter that needs careful monitoring to ensure the quality and consistency of the final biopharmaceutical drug product. The analytical method of choice used to perform this task is size-exclusion chromatography (SEC). However, it is becoming more and more apparent that considerable care is required in assessing the accuracy of SEC data. One old analytical tool that is now reappearing to help in this assessment is analytical ultracentrifugation (AUC). Developments in AUC hardware and, more importantly, recent developments in AUC data analysis computer programs have converged to provide this old biophysical tool with the ability to extract very high resolution size information about the molecules in a given sample from a simple sedimentation velocity experiment. In addition, AUC allows sample testing to be conducted in the exact or nearly exact liquid formulation or reconstituted liquid formulation of the biopharmaceutical in the vial, with minimal surface area contact with extraneous materials. As a result, AUC analysis can provide detailed information on the aggregation of a biopharmaceutical, while avoiding many of the major problems that can plague SEC, thus allowing AUC to be used as an orthogonal method to verity SEC aggregation information and the associating properties of biopharmaceuticals.
Authors and Affiliations
Steven A. Berkowitz
High-Throughput screeening assays for CYP2B6 metabolism and inhibition usuing fluorogenic vivid substrates
CYP2B6 is a highly polymorphic P450 isozyme involved in the metabolism of endo-and xenobiotics with known implications for the activation of many procarcinogens resulting in carcinogenesis. However, lack of validated hig...
G.L. Amidon, H. Lennernas, V.P. Shah, and J.R. Crison. A Theoretical Basis for a Biopharmaceutic Drug Classification: The Correlation of In Vitro Drug Product Dissolution and In Vivo Bioavailability, Pharm Res 12, 413–420, 1995—Backstory of BCS
The Biopharmaceutics Classification System (BCS) has become widely accepted today in the academic, industrial, and regulatory world. While the initial application of the BCS was to regulatory science bioequivalence (BE)...
Safety and Biocompatibility of Carbohydrate-Functionalized Polyanhydride Nanoparticles
Carbohydrate functionalization of nanoparticles allows for targeting of C-type lectin receptors. This family of pattern recognition receptors expressed on innate immune cells, such as macrophages and dendritic cells, can...
Ligand-Binding Assay Development: What Do You Want to Measure Versus What You Are Measuring?
The analysis of biotherapeutics by ligand-binding assay (LBA) is associated with some unique challenges that are unlike those commonly encountered in chromatographic methods for chemically based small molecule drugs. Whi...
Statistical Considerations for Assessment of Bioanalytical Incurred Sample Reproducibility
Bioanalytical method validation is generally conducted using standards and quality control (QC) samples which are prepared to be as similar as possible to the study samples (incurred samples) which are to be analyzed. Ho...