Automated high throughput protein stability measurement system

"To ensure product safety and efficacy, protein therapeutic critical quality attributes must remained within tolerances for the duration of the shelf life of the product, as well as during transportation, storage and use. Consequently development of and testing for long term stability of the protein drug product is a major focus of biotherapeutic development and formulation activities.

Thermodynamic stability of the higher order structure of the protein is used as a proxy for longer term stability of the drug product, allowing a large number of potential formulation conditions or variants on the drug product to be screened in a period of time significantly shorter than the potential shelf life, greatly increasing the speed the drug can be developed and commercialised.

The tools currently deployed are Differential Scanning Calorimetry (DSC), that looks at the change in heat capacity for the protein as it is heated, and the optical spectroscopic methods that look at changes in the higher order structure (HOS) of the protein with temperature. These approaches have traditionally been restricted by low throughputs (1 sample per hour) and high sample consumption.

Another approach repurposes qPCR machines by adding fluorescent dyes, that change their fluorescence properties on binding to the exposed hydrophobic core of the protein as it unfolds during heating. This approach allows screening of many conditions at once, in 384 well disposable microplates at relatively little cost per sample. But the utilisation of hydrophobic dye can produce false results due to other binding modes of the intact protein and interference with the dye binding from other buffer components.

This project is a feasibility study to determine the technical and commercial viability of a new high throughput technology for the screening of protein stability that requires very little sample, is able to be extensively automated and does not require addition of an indicating dye. The target market will be biopharmaceutical development and formulation, allowing much higher numbers of drug candidate variants or formulation conditions to be screened earlier in the development cycle."