5mg Soluble Protein Protein Refolding Protein Expression And Purification

Introduction

E.coli is the preferential one of bacterial systems for recombinant protein production and large scale fermentation as its advantages including fast rate of reproduction, ease of culture, and rich knowledge about its genetics. Bacterial protein expression service offered by CELLFREE provide a relatively simple, fast, and inexpensive platform for native and heterologous recombinant protein production.

Competitive Advantages

• Increase the protein expression level in most host cells
• Optimized expression vectors designed to improve the expression level
• Deliver 3-5 mg protein of 85% purity
• Streamlined process from gene synthesis and codon optimization to protein expression and purification

Service Content 

Service Process

• Codon Optimization

The eukaryotic protein expressed in E. coli may form the inactive aggregates of protein known as inclusion bodies. Using our unique bio-informatics software MaxcodonTM, we can reduce the chance of the formation of inclusion bodies in expression process.

• Washing

If the expressed recombinant proteins aggregate into the form of inclusion bodies, the first step of refolding is washing as inclusion bodies contains some outer membrane proteins, plasmid DNA, etc. The washing buffer is usually carried out by using a lower concentration of denaturant (such as urea) or a mild detergent (such as Triton X-100).

• Dissolution

After washing, inclusion bodies need dissolute for refolding. Denaturing agents such as urea and guanidine hydrochloride can break through various chemical bonds between the molecules of inclusion bodies, resulting in dissolution of inclusion bodies.

• Refolding

Generally, the refolding methods mainly include dilution, dialysis, molecular sieve chromatography and ion exchange chromatography. Four different methods have their own advantages and disadvantages. Please contact us for more detailed information.