A REVIEW: AGGREGATION ANALYSIS OF THERAPEUTIC PROTEINS
*Chilka R Patel, Dhara H Niranjani, Kirangi B Desai, P. K. Pradhan, U. M. Upadhyay
Department of Quality Assurance, Sigma Institute of Pharmacy, Vadodara, Gujarat, India.
ABSTRACT
Control and analysis of protein aggregation is an increasing challenge
to pharmaceutical research and development. Due to the nature of
protein interactions, protein aggregation may occur at various points
throughout the lifetime of a protein and may be of different quantity
and quality such as size, shape, morphology. It is therefore important
to understand the interactions, causes and analyses of such aggregates
in order to control protein aggregation to enable successful products.
This review gives a short outline of currently discussed pathways and
induction methods for protein aggregation and describes currently
employed set of analytical techniques and emerging technologies for
aggregate detection, characterization and quantification. A major
challenge for the analysis of protein aggregates is that no single
analytical method exists to cover the entire size range or type of
aggregates which may appear. Each analytical method not only shows
its specific advantages but also has its limitations. Size-exclusion chromatography (SEC) has
been a workhorse for detecting and quantifying protein aggregation ([1,2]). But SEC is often
questioned because of possible loss of proteins (soluble aggregates, in particular) by their
nonspecific binding to the columns ([3]). Native gels have also been used to observe protein
aggregation, but only qualitatively. Column-free techniques such as analytical
ultracentrifugation (AUC), field-flow fractionation (FFF), and dynamic light scattering (DLS)
now find increasing application in aggregation analysis.
Keywords: Aggregation, Electrophoresis, Size-Exclusion Chromatography,Analytical Ultracentrifugation, Dynamic Light Scattering, Field-Flow Fractionation.
[Full Text Article]