Information-rich, label-free spectroscopy as a tool for testing the stability of biotherapeutic formulations

D. Gregson and L. Cole, Applied Photophysics Ltd, 201-205 Kingston Road, Leatherhead KT22 7PB, United Kingdom

Abstract

A stable, correctly-folded protein is an absolute requirement for a successful biotherapeutic and the stability of a desired conformation is determined by suitable formulation.

Applied Photophysics has developed a high-content, label-free technique, known as dynamic multi-mode spectroscopy (DMS), to study the thermal stability of proteins in solution. As an example we report an investigation into the stability of a monoclonal antibody biotherapeutic in different formulation conditions, using DMS.

An interest had been expressed in discovering whether DMS could dentify any differences in the antibody denaturation signatures in acetate and lactate formulations, following a request by the regulatory authorities to explain an apparent difference in T_m that had been highlighted by differential scanning calorimetry (DSC). The DSC experiments showed that the final transition of the biotherapeutic formulated in lactate exhibited a T_m that was 2°C higher than it was when formulated in acetate. DMS is well-suited to address such questions because it generates both structural and thermodynamic data together with information on aggregation, in a single experiment. The samples were measured over a period of weeks and it became apparent that the antibody behaved differently as it aged in the two formulations, with the lactate formulation imparting greater robustness than the acetate.

The study gives insights into the sample behaviour that might not be readily apparent using one-dimensional techniques, demonstrating the advantages of generating both structural and thermodynamic information in a single, sample-efficient measurement.

About Applied Photophysics

Applied Photophysics is a world-leading manufacturer and supplier of precision spectrometers for the study of circular dichroism, stopped-flow kinetics and laser flash photolysis.

Applied Photophysics Limited was established in 1971 by The Royal Institution of Great Britain.

 

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