Cell free protein expression is increasingly becoming a critical tool across workflows such as enzyme engineering, synthetic biology, biocatalysis development and diagnostic reagent production. While conventional cell-based expression systems (e.g. E. coli, yeast, or mammalian hosts) remain widely used, they often impose biological constraints that limit their suitability for complex or difficult protein targets.
Limitations of Cell-based Systems
Common challenges typically encountered with cell-based systems include:
- Prolonged expression times due to cellular growth cycles
- Poor expression yields for toxic or unstable proteins
- Difficulty producing membrane proteins or multi-domain complexes
- Low throughput for screening large variant libraries
- Proteolytic degradation
As a result, when rapid design-build-test cycles or incompatible targets are involved, traditional expression systems become a bottleneck. In these situations, cell-free protein synthesis (CFPS) offers a powerful alternative.
ProteoSynth®: Cell Free Protein Synthesis Platform
To address these limitations, ProteoSynth® is a proprietary cell-free protein synthesis platform developed by Isomerase to address the limitations of cell-based expression systems. Unlike conventional approaches, CFPS decouples protein production from living cells by isolating the translational machinery (ribosomes, tRNAs, enzymes, cofactors) directly from E. coli BL21(DE3). Consequently, this provides unprecedented control over expression conditions.
By removing cellular viability constraints, ProteoSynth® enables researchers to:
- Express cytotoxic or aggregation-prone proteins
- Produce proteins/peptides incorporating modified or non-canonical amino acids
- Avoid membrane transport and in vivo folding limitations
- Operate in highly controlled reaction environments free of nucleases and proteases
Moreover, in the ProteoSynth® system, transcription is driven by T7 RNA polymerase, and translation occurs under carefully optimised salt, energy and cofactor conditions. As a result, this setup enables teams to express proteins within hours, offering a highly responsive platform for demanding applications.
Key Advantages of Cell Free Protein Synthesis with ProteoSynth®
- Achieve rapid expression timelines: from DNA template to protein in hours
- Access high compatibility: supports a wide range of protein classes
- Overcome toxicity constraints: enables production of proteins that exhibit cytotoxic effects on host cells in vivo
- Eliminate enzymatic degradation: protein production in the absence of nucleases or proteases
- Integrate seamlessly: fits into high-throughput protein engineering pipelines
- Assemble synthetic cascade or multi-enzyme pathways rapidly in vitro, allowing iterative optimisation of multi-step biocatalytic routes before full-scale development
- Accelerate screening workflows: conduct microscale synthesis for kinetic studies, substrate screening and reaction optimisation
Therefore, ProteoSynth® empowers both discovery and development teams to compress timelines and overcome expression barriers that limit conventional systems.
Flexible Access Models to Fit Your Project Needs
Service Model
Isomerase offers CFPS as a service for clients requiring protein production without installing in-house workflows:
- Partner provides DNA sequences
- We conduct codon optimisation
- We evaluate expression yield, solubility and activity
- Analytical data provided
- Protein is supplied at requested purity and quantity
Standalone Kit
Our CFPS is available as a deployable system for partners seeking to operate CFPS workflows directly.
- Delivered as lysate
- Suitable for variant library screening and prototyping
Common Application of Cell Free Protein Expression
- Directed evolution campaigns (library expression & variant screening)
- Expression of toxic or aggregation-prone proteins
- Peptide synthesis
- Membrane protein production
- Biocatalyst development
Summary
In summary, ProteoSynth® delivers the full power of cell-free protein synthesis to enable rapid, flexible, and scalable protein production, particularly for targets where conventional systems fail. Contact our team to explore how ProteoSynth® can support your protein expression goals.