Introducing ALiCE® - Simple, scalable, cell-free expression of difficult-to-express proteins with Live Q&A

Many commercially promising proteins are difficult-to-express (DTE) in living cells and are lost before an industrial production process can be established. Cell-free protein expression (CFPE) is well suited to producing DTE proteins but has largely been limited to microgram yields.

Here we present a eukaryotic lysate, sold as ALiCE® by LenioBio GmbH, which produces protein yields of up to 3mg/ml within a 48 hour reaction time. Being a eukaryotic system, ALiCE® has incredible promise for protein engineering due to its capabilities in performing post-translational modifications including disulfide bonds and N-glycosylation.

Recent developments in the scaling of ALiCE® bioprocessing now permit the production of liters of lysate, presenting a new challenge in scaling of the CFPE reaction to match.

We present case studies of cytosolic and a microsomally expressed proteins produced by ALiCE®, and describe their subsequent purification and characterization. We further disclose world first liter scale eukaryotic CFPE reactions using commercially available bioreactors.

Learning Objectives:

1. Discuss the difficulties of producing certain protein classes.

2. Explain why cell-free protein expression technology is more appropriate for producing certain classes of proteins.

3. Discuss factors important in scaling of cell-free protein expression for commercial scale protein manufacturing.