New work could provide sustainable ways to make chemicals, medicines, and biomaterials. Single-pot Glycoprotein Biosynthesis Using a Cell-Free Transcription-Translation System Enriched with Glycosylation Machinery, the study was published in Nature Communications.
Cell-Free Protein Synthesis
Engineering cellular biology, minus the actual cell, is a growing area of interest in biotechnology and synthetic biology. It's known as cell-free protein synthesis or CFPS, and it has the potential to provide sustainable ways to make chemicals, medicines, and biomaterials.
Unfortunately, a long-standing gap in cell-free systems is the ability to manufacture glycosylated proteins with a carbohydrate attachment. Glycosylation is crucial for a wide range of important biological processes, and the ability to understand and control this mechanism is vital for disease treatment and prevention.
They have teamed up on a novel approach that bridges this gap. Their system, the first of its kind, capitalizes on the recent advances in CFPS while adding the crucial glycosylation component in a simplified, "one-pot" reaction. The protein of choice could then be freeze-dried and reactivated for point-of-use synthesis by simply adding water.
This work could impact the development of decentralized manufacturing strategies. Rapid access to protein-based medicines in remote settings could change lives; new biomanufacturing paradigms suitable for use in low-resource settings might promote better access to costly drugs through local, small-batch production.
For their new method, the team prepared cell extracts from an optimized laboratory strain of E. coli, CLM24, that were selectively enriched with key glycosylation components. The resulting extracts enabled a simplified reaction scheme, which the team has dubbed cell-free glycoprotein synthesis (CFGpS).
A major advance of this work is that our cell-free extracts contain all of the molecular machinery for protein synthesis and protein glycosylation. What that means is you only need to add DNA instructions for your protein of interest to make a glycoprotein in CFGpS. This is a drastic simplification from cell-based methods and allows us to make sophisticated glycoprotein molecules in less than a day.
And the CFGpS method is highly modular, allowing for the use of distinct and diverse extracts to be mixed for the production of a variety of glycoproteins. Because we chose E. coli, which lacks its glycosylation machinery, to build our CFGpS platform, it gave us a blank slate for bottom-up engineering of any desired glycosylation system.
This gives us the unique ability to control carbohydrate structures and purities of the glycoproteins at levels that are not currently achievable in other cell-based expression systems. Even in developed countries like the U.S., the move toward personalized medicine makes this type of on-demand drug production protocol attractive.