Use of Microcalorimetry and Its Correlation with Size Exclusion Chromatography for Rapid Screening of the Physical Stability of Large Pharmaceutical Proteins in Solution

Abstract

The utility of microcalorimetry as a rapid screening tool for assessing the solution stability of high molecular weight pharmaceutical proteins was evaluated by using model recombinant antibodies, Protein I and Protein II. Changes in the transition midpoint, T(m), were monitored as a function of pH and/or in the presence of excipients, and results were compared with traditional accelerated stability data from samples that were analyzed by size exclusion chromatography (SEC). The data from microcalorimetry were well correlated with those from SEC for predicting both optimal solution pH as well as excipient effects on solution stability. These results indicate that microcalorimetry can be an efficient screening tool useful in identifying optimal pH conditions and excipients to stabilize pharmaceutical proteins in solution formulations.

Opposite Effects of Polyols on Antibody Aggregation: Thermal Versus Mechanical Stresses

“…Attempts have been made to correlate T m of proteins in different solution conditions with the trend in aggregation observed upon high temperature incubation of proteins (11,12). Since both, DSC and incubation, involve subjecting proteins to thermal stress, it is likely that the observed correlation arises from unfolding in the bulk solution being the governing mechanism of aggregation in these studies.…”

Biophysical and formulation studies of theSchistosoma mansoniTSP-2 extracellular domain recombinant protein, a lead vaccine candidate antigen for intestinal schistosomiasis

“…Compounds were defined as having a stabilizing effect when they increased the thermal melting temperature (Tm) values of the first structural transition, as seen in the empirical phase diagram of the protein, which is often correlated with storage stability. 26 Screening employed a library of GRAS compounds. The conditions of the screening were determined by inspection of the apparent phase boundaries established by the EPD.…”

Correlation Between Enzyme Activity and Stability of a Protease, an Alpha‐Amylase and a Lipase in a Simplified Liquid Laundry Detergent System, Determined by Differential Scanning Calorimetry

“…Residual activity after 14 days at 35°C was used to create the stacked plots (lines connecting the data are for guidance of the eye only) (2). The gray squares were omitted from the regression analysis (see text) [73][74][75][76], but in situations where the unfolding is either not kinetically controlled, in the solid state, or where thermal stability is compared to size-exclusion chromatography (SEC) data. In the latter case, aggregation is the irreversible end-point of the denaturation.…”