Predicting the physical stability of biopolymers by self-interaction chromatography
Date
2008
Journal Title
Journal ISSN
Volume Title
Abstract
Biopolymers, including proteins and peptides, are increasingly taking the place of small molecules in manufacturing, health and home applications. The advantage of using biopolymers over small molecules for therapeutic applications include high activity, high specificity and low toxicity. The disadvantages of using biopolymers include large scale production, chemical instability and physical degradation. A biopolymer that shows promise for a specific application will not advance past early development if the protein lacks either chemical or physical stability. Of these two parameters, physical stability is often harder to predict and/or measure. The physical stability of a biopolymer can be improved by site-directed mutagenesis, glycosylation, post-translational modification and adjusting the solvent/co-solvent system. Quantitatively measuring the change in the physical stability of a protein in different solvent/co-solvent systems can increase the number of early developmental stage proteins that advance to commercial scale. Physical stability of proteins can be described by protein-protein interactions and measured by the osmotic second virial coefficient (B). However, current methods used to measure B such as static light scattering are not practical for large screening studies because of large protein consumption, low throughput, method variability and analyte size limitations. An alternative method to measure B is Self-Interaction Chromatography (SIC), which requires less protein, shorter analysis time, allows for miniaturization and capable of measuring B for small size biopolymer. The ability of SIC to measure B for a therapeutic protein, a peptide and membrane proteins in different solvent/co-solvent systems has been demonstrated in this dissertation.
Description
Rights Access
Subject
biopolymers
HPLC
peptides
proteins
self-interaction chromatography
stability
analytical chemistry