Expression, in vivo Maturation And Tag-Less Purification Of A Recombinant Protein

To transition to a cost-effective and scalable process, the existing workflow must be redesigned, with a focus on recombinantly expressing the untagged protein in E. coli and facilitating in vivo maturation. The first critical step in this process is optimizing expression parameters to ensure the mature protein is enriched in the soluble cytoplasmic fraction. Once this is achieved, a scalable purification strategy must be developed to meet both the quality and quantity specifications for the final protein product.

This case study highlights the expertise and experience of Aragen’s scientists in assisting a client in re-engineering their production strategy to reach a critical milestone. The client aimed to obtain more than 50 mg of a mammalian secretory protein, which is notoriously difficult to express as a soluble form in E. coli. Aragen was approached to recombinantly produce the protein in its precursor form, facilitate in vivo maturation through co-expression with a processing enzyme, and purify the tag-less protein to greater than 95% purity. This project was pivotal for the client, enabling the initiation of a new program in their drug discovery pipeline and laying the foundation for further development.