Cell-Free Protein Expression
High-throughput protein production using cell-free expression systems for rapid prototyping and functional studies
Cell-free protein expression is a revolutionary approach to protein production that eliminates the need for living cells. Our platform utilizes optimized cell-free systems to produce high-quality proteins rapidly and efficiently.
How It Works
Cell-free protein expression systems contain all the necessary components for protein synthesis—ribosomes, tRNAs, amino acids, and energy sources—extracted from cells and reconstituted in a controlled environment.
DNA Template Preparation
Your protein sequences are cloned into optimized expression vectors with strong promoters and ribosome binding sites.
Our vectors are pre-optimized for cell-free systems, eliminating the need for complex cloning optimization.
Cell-Free Reaction Setup
DNA templates are added to cell-free lysates containing all necessary translation machinery.
We use wheat germ and E. coli-based cell-free systems depending on your protein requirements.
Protein Synthesis
Proteins are synthesized in a controlled environment at optimal temperature and conditions.
Typical expression times range from 2-16 hours depending on protein complexity.
Key Advantages
Rapid Prototyping
- Fast turnaround: Proteins expressed within hours, not days
- High-throughput: Multiple variants processed simultaneously
- No cell culture: Eliminates growth phases and maintenance
Flexibility
- Toxic proteins: Express proteins that would be lethal to living cells
- Membrane proteins: Specialized systems for challenging targets
- Post-translational modifications: Controlled modification environments
Quality Control
- Consistent conditions: Reproducible expression across batches
- Real-time monitoring: Track expression progress in real-time
- Immediate analysis: Direct compatibility with downstream assays
Applications
Protein Screening
Rapid expression of protein variants for functional screening and characterization.
Binding Studies
Direct expression for immediate use in binding assays and kinetic studies.
Enzyme Assays
Fresh enzyme production for activity measurements and optimization.
Structural Studies
High-quality protein production for crystallography and NMR studies.
Expression Systems
Best for: Cytoplasmic proteins, enzymes, binding domains
Features:
- High expression levels
- Cost-effective
- Excellent for prokaryotic proteins
- Compatible with isotope labeling
Typical yield: 0.1-2 mg/mL
Best for: Cytoplasmic proteins, enzymes, binding domains
Features:
- High expression levels
- Cost-effective
- Excellent for prokaryotic proteins
- Compatible with isotope labeling
Typical yield: 0.1-2 mg/mL
Best for: Eukaryotic proteins, membrane proteins, complex domains
Features:
- Eukaryotic protein folding machinery
- Disulfide bond formation
- Reduced proteolysis
- Compatible with eukaryotic modifications
Typical yield: 0.05-0.5 mg/mL
Best for: Challenging proteins, membrane proteins, glycoproteins
Features:
- Optimized for specific protein classes
- Enhanced folding environments
- Customizable cofactor addition
- Specialized for difficult targets
Typical yield: Variable, optimized per target
Quality Metrics
Our cell-free expression systems are monitored for:
Integration with Other Technologies
Cell-free expression seamlessly integrates with our other platform technologies:
- Affinity purification: Direct capture using tagged proteins
- Binding assays: Immediate use in BLI and SPR studies
- Quantification: Real-time monitoring with split-GFP systems
- Characterization: Direct input to nanoDSF and HPLC-SEC analysis
While cell-free systems offer rapid protein production, some proteins may require optimization for proper folding and activity. Our team provides guidance on system selection and optimization strategies.