Cell-free Expression System

Introduction

The so-called non-cell protein expression system refers to the expression of the protein by using in vitro translation system (such as ribosome display technology) to mimic the protein translation system in cell, at the same time using the well-designed liposome membrane to replace the cell membrane, proteins were inserted on it to maintain the maximum natural conformation and natural activity.

In vitro protein expression is the production of recombinant proteins in solution using biomolecular translation machinery extracted from cells. Because protein synthesis occurs in cell lysates rather than within cultured cells, the method is also called cell-free protein expression. Cell-free protein production can be accomplished with several kinds and species of cell extract, and these approaches have several advantages and features that complement traditional in vivo methods.

Introduction to in vitro protein expression

In vitro protein expression (also known as in vitro translation, cell-free protein expression, cell-free translation, or cell-free protein synthesis) is a technique that enables researchers to rapidly express and manufacture small amounts of functional proteins. Compared to in vivo techniques based on bacterial or tissue culture cells, in vitro protein expression is considerably faster because it does not require gene transfection, cell culture or extensive protein purification.

Although in vitro expression is not practical for commercial large-scale recombinant protein production, it has a variety of features that make it considerably more useful and flexible for many research applications.

Applications for cell-free protein expression

• Experiments to characterize protein-protein interactions and protein-nucleic acid interactions
• Rapid and high-throughput expression of mutant or truncated proteins for functional analysis
• Expression of mammalian proteins with proper glycosylation and native post-translational modifications (PTMs)
• Labeling of proteins with stable isotopes for structural analysis
• Production of functional virons or toxic polypeptides
• Analysis of components required for protein folding, protein stability or protein degradation

The diagram provides an example of the application of a cell-free protein expression method combined with protein mass spectrometry (MS).

Production of cell-free membrane proteins

• Expression of membrane protein in bacteria;
• Expression of membrane protein in wheat embryo;
• Expression of membrane protein in Baculovirus;
• Expression of membrane protein in rabbit reticulocyte.

Advantages

• Proprietary Technologies
• Guarantees your protein expression and purification
• Competitive price
• Fast turnaround: as little as 4 weeks
• Flexible scale-up protein production

Comparison of various cell-free protein expression systems

System

Advantages

Disadvantages

E. coli

· Very high protein yield

· Relatively tolerant of additives

· Many eukaryotic proteins insoluble upon expression

· Eukaryotic co- and post-translational modifications not possible

· Codon usage is different from eukaryotes

Rabbit

· Mammalian system

· Cap independent translation

· Sensitive to additives

· Protein glycosylation not possible

· Co-expression of off-target proteins

Wheat germ

· Translation of large proteins possible

· Devoid of off-target endogenous mammalian proteins

· High protein yield

· Mammalian co- and post-translational modifications are not possible

· Premature termination of products

Insect

· Translation of large proteins possible

· No endogenous mammalian proteins

· Certain forms of protein glycosylation possible

· Non-mammalian

Human

· Human system

· Co- and post-translational modifications are possible

· Synthesis of functional proteins

· Possible to make virus-like particles (VPLs)

· Sensitive to additives

· Lower yields than E. coli

· New system

Related Services

Protein Expression in Bacteria

Unique Protein Production Service

Protein Expression in Cell-Free System

Rich in Disulfide Bond Protein Expression

Protein Expression in Mammalian Cells

Protein Expression in Baculovirus

Protein Expression in Yeast

Protein Expression in Bacillus subtilis

Protein Expression in E coli

Virus-like particles/Phage library expression system based

Cell-free expression system

Whole cell-based protein expression system

Why Leading Biology?

At Leading Biology, we custom protein purification design for every single protein to ensure the production and recovery rate as high as possible.

Working with us, you will get stability, and it means a reliable partner to help streamline your R&D process.

Working with us, you will get the guaranteed service to accommodate your requirements.

• Innovative configurations of chromatography columns custom tailored to each protein
• Vigorous quality control system to ensure the required quality and reproducibility
• Production capacity of up to tens of grams
• Flexible scale-up protein production
• Competitive price with fast turnaround time

Contact Information

Please obtain a quote before ordering, and refer to the quote number when you place an order.

Orders are typically confirmed within 12 hours.

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