TYPICAL CASE STUDY:

A client approached us with a laboratory scale process for the purification of a recombinant protein product which was produced by an Escherichia coli strain. Following technology transfer of the clients procedures into Angel Biotechnology, the process was scaled up and a cGMP compliant process was established. Several of the process steps were known to be sub-optimal and these required further development.

• The production process was scaled up from a 1L shake flask culture to a 50L cGMP compliant fermentation
• The original culture medium contained an animal-derived component which was replaced with a similar component derived from a non-animal source
• The original cell harvesting procedure relied on batch centrifugation. A tangential flow filtration process was established as a more readily scaleable alternative
• The cell lysis procedure required sonication coupled with the addition of expensive lysozyme/DNAase enzyme reagents to the cell suspension which was kept on ice to avoid overheating. This procedure was considered impractical to scale up in a cGMP environment with the added disadvantage of introducing potential process contaminants and so it was replaced with a highly efficient fully automated mechanical homogenization process
• The original refolding procedure involved a dialysis procedure which was difficult and impractical to scale up. The procedure was replaced with a procedure where the refolding temperature was tightly controlled and which could be easily scaled up
• Protein chromatography procedures were established using an AktaTM/Unicorn based platform
• A comprehensive set of analytical procedures were developed. These were then used to analyse in-process and final product samples for product and process related impurities and contaminants. One of the analytical methods we developed identified a product-related impurity in the original manufacturing process. This was further characterized using mass spectrometry