Cell-Free Expression Technology

Cell-free expression systems have become a mature technology for synthesizing proteins in cell lysates. CD Formulation integrates cell-free expression technology into our internal protein production platform, offering the ultimate solution for your protein production. Our services can be tailored to your specific requirements (e.g., tag selection, purity).

What is The Cell-Free Expression Technology?

Cell-free protein expression, also known as in vitro translation, cell-free translation, and cell-free protein synthesis, is a method of synthesizing proteins in vitro that can produce target proteins without the use of living cells. Because this protein synthesis occurs in cell lysates rather than in cultured cells, this method is called cell-free. This technology enables researchers to quickly express and manufacture small amounts of functional proteins. Compared to in vivo techniques in bacteria or tissue culture cells, cell-free protein expression is fast because no gene transfection, cell culture, or extensive protein purification is required.

Fig. 1 The components of a cell-free protein synthesis reaction. (Brookwell A, et al., 2021)

Our Services Related to Cell-Free Expression Technology

CD Formulation is one of the world leaders in the production of therapeutic proteins using advanced cell-free protein synthesis technology. Thanks to the innovative cell-free protein synthesis system (CFPS), we can handle difficult proteins that show no/low expression, aggregation, etc. in other systems. The expression process that originally took several days has been optimized and shortened to a few hours.

Our personalized cell-free protein synthesis service can flexibly express proteins according to the specific needs of our customers. By introducing the desired nucleotide modifications in the transcribed tRNA, we can help increase the freedom of regulation of the translation process.

Our services include but are not limited to:

Gene Synthesis and Codon Optimization (Optional)

• Start with the antibody sequence

Vector Construction (Optional)

• Cloning into an expression vector
• Plasmid sequencing
• Plasmid generation
• Cell-free expression and purification

Quality Control (QC)

• SDS-PAGE
• UV
• SEC-HPLC

Deliverables

• Purified protein

Key Components and Processes of Cell-Free Expression Technology

Two basic components of cell-free protein expression are a genetic template (plasmid DNA, linear DNA, or mRNA) encoding the protein of interest and a reaction solution containing the necessary molecular machinery for transcription and translation.

In CFPS, our scientists use a solution containing all the cellular machinery required to direct protein synthesis to transcribe and translate the provided nucleic acid template.

Common cell extracts contain most or all of the molecules in the reaction solution, including:

• RNA polymerase for mRNA transcription.
• Ribosomes for polypeptide translation.
• tRNA and amino acids.
• Enzyme cofactors and energy sources.
• Necessary cellular components for correct protein folding.

Different Cell-Free Expression Systems

Extracts from cell-free expression are made using different systems to support high levels of protein synthesis. The following table lists the common cell-free expression systems.

Custom Cell-Free Expression Services

Features of Cell-Free Expression Technology

• Cell-free reactions are easy to set up and require only a few hours to express proteins.
• Using small-scale reactions (50-100uL), reaction conditions can be optimized quickly and in parallel.
• Milligram quantities of soluble protein can be obtained from a few milliliters of reaction volume.
• Eukaryotic cell-free lysates provide post-translational modifications such as core glycosylation or phosphorylation and native membrane components for membrane protein insertion.
• Proteins and complexes with multimeric conformations can be expressed.
• The open system allows the addition of many components to the reaction, such as molecular chaperones, labeled xenyl acids, or detergents, allowing for direct manipulation of the chemical environment.

Publication

Thanks to our in-house advanced cell-free expression platform, CD Formulation is committed to providing efficient and economical production of therapeutic proteins. Please feel free to contact us if you are interested in our services. Learn how our cell-free expression technology can support your recombinant protein synthesis projects.  

How It Works

STEP 1

Submit a service request describing your specific research or development need.

STEP 2

We'll email you to provide your quote and confirm order details if applicable.

STEP 3

Execute the project with real-time communication, and deliver the final report promptly.