BioMarin Pharmaceutical -Donnie Mackenzie

A critical step in a protein’s purification process is to identify formulation buffers for optimal stability and activity. Before the Prometheus, we were primarily using a mixture of spectrophotometry, SDS-PAGE, size exclusion chromatography, multi-angle/dynamic light scattering, and various dyes used for aggregation and thermal shift assays. These methods gave us a stitched together picture of protein stability. However, each assay had its limitations. The more we manipulated the proteins to accommodate the compatibilities of the assays the less confidence we had that we were fully optimizing their stability. We were also sacrificing large amounts of our hard-fought purified material. Now with the Prometheus, we can get away with using 10uL of nanomolar concentrations of material per data point and acquire stability information all in one assay. We have been able to save on reagents, time, and costs with the throughput of this instrument. We have expanded the types and amounts of formulations we can test including denaturants, high salts, and detergents which are tricky for other methods. The Prometheus enabled us to identify stabilizing agents for complex proteins with multiple domains. The Prometheus technology is also being utilized for drug discovery efforts to identify small molecule binding to target domains. We continue challenging the instrument with complicated proteins and formulations; and it keeps giving us reproducible, high-quality data that allows us to get high quality therapeutics to patients faster.


 

Donnie Mackenzie
Scientist II
BioMarin Pharmaceutical