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The cell-free protein expression system, also referred to as the in vitro translation system, utilizes target mRNA or DNA as a template. Substrates and energy essential for protein synthesis are added to the enzyme system derived from cell extract, enabling the synthesis of target proteins in vitro. This system mimics in vivo cells, replicating the intracellular processes of protein transcription and translation. Various materials necessary for protein synthesis, such as energy, transcription factors, and translation factors, are required for its operation.
This system is especially well-suited for expressing transmembrane and toxic proteins, known for its short processing cycle and high throughput.
Service Process
1. Plasmid construction and preparation
Process
- Codon optimisation and gene synthesis
- The PCR amplification products undergo restriction enzyme digestion before being ligated to the pET vectors
- Large quantities of recombinant plasmids are generated
Features
Multi-vector optimization - To enhance mRNA translation efficiency and protein yield, we offer a protein expression service that optimizes N-terminal peptides alongside conventional fusion proteins. These peptides, 6-11 amino acids long, represent our shortest custom-designed sequences for this purpose.
Lead Time: 15-20 Business Days
2. Small-scale expression and optimisation
Process
- Optimization across multiple conditions
- Gel electrophoresis using SDS-PAGE
- Identifying the ideal reaction parameters
Features
In a cell-free expression system, multiple proteins can be simultaneously expressed on a multi-well plate across various conditions. This enables us to provide a service for optimising conditions across multiple parameters.
Lead Time: 10-15 Business Days
3. Target protein expression and purification
Process
The target protein undergoes purification by exploring various chromatographic conditions, such as ion exchange chromatography, size exclusion chromatography, and others, utilizing AKTA equipment. Subsequently, the optimal purification method is determined.
Features
Multi-condition purification scheme (optional) - For transmembrane proteins, we offer diverse detergent purification services to optimise your protein's buffer. Ideal for bioactive transmembrane proteins.
Lead Time: Additional 3 Business Days for Multi-Condition Purification
4. Optional additional services
Process
- Endotoxin removal
- Lyophilisation
- Filter sterilisation
- Tag removal (Additional charge)
Features
Customers have the flexibility to select from a range of additional services tailored to their specific requirements.
Lead Time: 3-5 Business Days
5. Quality control
Process
The purity, concentration and further tests are performed.
Features
Detailed COA report is provided.