Trusted by Leading Research & Pharma Institutions

E. coli Cell-Free Protein Synthesis

Rapid, high-yield in vitro protein expression without the constraints of living cells. Synthesize functional proteins in hours instead of days using our optimized S30 extract platform.

T7 Promoter Control

Hours to Results

0.5+ mg/mL Yield

ISO Quality

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Trusted by leading research and pharmaceutical institutions

MIT

Pfizer

Stanford

Roche

Johns Hopkins

Merck

Why Choose Us

Gene to purified protein in 1-3 days

Express toxic proteins successfully

Flexible engineering options

Scalable from 50 uL to liters

Rapid Expression

1-3 business days from gene to protein

Toxic Protein Friendly

Express proteins lethal to living cells

Flexible Engineering

Non-canonical amino acids and modifications

Yield

0.5+ mg/mL

Service Overview

Why Choose Our E. coli CFPS Service

Advanced cell-free protein synthesis platform delivering rapid, high-quality protein expression

Rapid Expression

Go from gene to purified protein in 1-3 business days. No cell culture, no colony screening, no waiting weeks for expression.

Toxic Protein Friendly

Express proteins that would kill or cripple living cells. Bypass metabolic constraints entirely with our open reaction system.

Flexible Engineering

Easy incorporation of non-canonical amino acids, labels, and modifications. Direct access to reaction components for experimental control.

Scalable Platform

From 50 uL screening reactions to multi-milligram production. Scale up without redesigning your workflow.

Quality Guaranteed

Every batch validated against control standards. Consistent yields and purity across multiple preparations with comprehensive QC.

Cost Effective

Eliminate expensive fermentation runs and downstream processing. Get more data from smaller budgets.

Technology Platform

Our CFPS Technology

Optimized platform technology for robust and reproducible protein expression

E. coli S30 Extract Platform

Our service utilizes optimized E. coli cell-free extract derived from engineered strains lacking RNase I. The extract contains all necessary transcription and translation machinery.

Endogenous TFs Ribosomes T7 Polymerase

Optimized Reaction Buffer

The reaction buffer contains precisely balanced components for maximum protein synthesis including amino acids, NTPs, and molecular crowding agents.

20 AA NTPs PEG-8000

Template Requirements

We accept various DNA template formats including plasmid DNA, linear DNA (PCR products), and mRNA for maximum experimental flexibility.

T7 Promoter RBS 5-15 kb

Quality Control

SDS Gel electrophoresis verification

WB Western blot confirmation

Activity Functional activity assay

MS Mass spectrometry verification

Purification Options

His His-tag affinity purification

GST GST-tag purification

MBP MBP-tag purification

None Untagged protein available

Specifications

Service Specifications

Comprehensive specifications for our E. coli CFPS platform

Parameter	Specification
Expression System	E. coli Cell-Free (S30 Extract)
Transcription	T7 RNA Polymerase (phage-derived)
Protein Size Range	10 - 230 kDa
Typical Yield	0.5 - 1.0 mg/mL
Reaction Volume	50 uL - 10 mL (customizable)
Template Format	Plasmid DNA, Linear DNA, mRNA
Incubation Temperature	16C - 37C (adjustable)
Incubation Time	4 - 24 hours
Quality Control	SDS-PAGE, Western Blot, Activity Assay
Delivery Format	Reaction mix, Purified protein, or Lyophilized

Workflow

Our Service Workflow

Streamlined five-step process from template submission to protein delivery

Template Submission

Submit your gene sequence in plasmid or digital format

Reaction Setup

Prepare cell-free reaction mix with optimized components

Expression

Incubation at optimal temperature for protein synthesis

Quality Analysis

SDS-PAGE, concentration measurement, and validation

Delivery

Shipped on dry ice with complete documentation

Applications

Diverse Applications Across Biotechnology

Our CFPS platform supports research and development in multiple fields

Research Applications

Our E. coli CFPS platform accelerates research workflows by eliminating the time-consuming steps of traditional expression systems. Get functional protein without cell transformation, colony screening, or fermentation optimization.

Functional protein expression and characterization
Protein-protein interaction studies
Enzyme kinetics and mechanism analysis
Epitope mapping and antibody validation

1-3 Days

Gene to purified protein

High-Throughput Screening

Parallel screening of hundreds of variants in microscale reactions dramatically accelerates protein engineering workflows. The cell-free format enables direct manipulation of reaction conditions without cell viability constraints.

Rapid variant library screening
Mutation impact analysis
Domain mapping experiments
Condition optimization studies

200+

Variants per batch

Protein Engineering

The open nature of the cell-free system provides unprecedented access to the synthesis machinery. Incorporate non-canonical amino acids, fluorescent labels, and other modifications directly into your protein of interest.

Non-canonical amino acid incorporation
Site-specific labeling (fluorescent, isotopic)
Fusion protein construction
Thermostability engineering

ncAA

Custom modifications

Testimonials

What Our Clients Say

Trusted by researchers worldwide for quality and reliability

"The cell-free system let us express a cytotoxic kinase domain that simply would not work in E. coli. We got functional protein in under a week."

Senior Researcher

Pharmaceutical Company

"We screened 200+ variants in parallel using microscale reactions. The speed difference compared to traditional expression is remarkable."

Principal Investigator

Academic Research Institution

"Incorporating unnatural amino acids for labeling was straightforward. The open nature of the system is a major advantage."

Postdoctoral Fellow

Biotechnology Company

Scientific Literature

Scientific Foundation

Our platform is backed by peer-reviewed research

85 Citations

Cell-Free PURE System: Evolution and Achievements

Cui Y, Chen X, Wang Z, Lu Y, et al. BioDesign Research. 2022.

Comprehensive review of PURE system elements, design, and applications in prototyping, non-natural protein synthesis, and biosensors.

View DOI

12 Citations

Adeno-Associated Virus 5 Protein Particles Produced by E. coli Cell-Free Protein Synthesis

Deuker D, Asilonu E, Bracewell DG, Frank S, et al. ACS Synthetic Biology. 2024.

Exploration of CFPS for producing AAV5-like virus-like particles, demonstrating successful expression and particle assembly.

View DOI

28 Citations

Complex Dependence of Escherichia coli-based Cell-Free Expression on Sonication Energy During Lysis

Piorino F, Styczynski MP, et al. ACS Synthetic Biology. 2023.

Investigation of sonication energy effects on cell-free protein synthesis productivity and optimization strategies.

View DOI

45 Citations

Activation of Energy Metabolism through Growth Media Reformulation Enables a 24-Hour Workflow for Cell-Free Expression

Levine MZ, So B, Mullin AC, et al. ACS Synthetic Biology. 2021.

Development of cell-free auto-induction medium enabling 24-hour workflow with sfGFP yields exceeding 1 mg/mL.

View DOI

52 Citations

Differentially Optimized Cell-Free Buffer Enables Robust Expression from Unprotected Linear DNA in Exonuclease-Deficient Extracts

Batista AC, Levrier A, Soudier P, et al. ACS Synthetic Biology. 2022.

Development of recBCD-deficient E. coli strains and optimized buffer enabling linear DNA expression without protection strategies.

View DOI

FAQ

Frequently Asked Questions

Find answers to common questions about our service

What types of proteins can be expressed in the E. coli CFPS system?

Our E. coli CFPS service can express a wide range of proteins from 10-230 kDa. This includes enzymes, fluorescent proteins, antibody fragments, transcription factors, and regulatory proteins. The system is particularly well-suited for proteins that are difficult to express in living cells due to toxicity, insolubility, or rapid degradation. While the system does not perform complex eukaryotic post-translational modifications, basic modifications like phosphorylation can occur.

Do I need a specific vector for expression?

Yes, your DNA template must contain a T7 promoter upstream of your gene of interest, along with a ribosome binding site (RBS) and a T7 terminator. Standard bacterial expression vectors like pET series work well. If you do not have a suitable vector, we can provide our optimized expression vectors as part of the service. Linear DNA templates (PCR products) are also accepted with appropriate preparation.

What yields can I expect from the cell-free reaction?

Typical yields range from 0.5-1.0 mg/mL of functional protein in standard batch reactions. The actual yield depends on the specific protein sequence, codon usage, and whether the protein tends to form inclusion bodies. For problematic proteins, we can optimize conditions by adjusting temperature, adding chaperones, or using fusion tags to improve solubility and yield.

Can you incorporate non-canonical amino acids?

Yes, our CFPS system supports incorporation of non-canonical amino acids (ncAA) for site-specific labeling or probing. Common applications include incorporation of fluorescent amino acids, isotopic labels for NMR, and photo-crosslinkers. We recommend consulting with our team to discuss your specific ncAA requirements and expected incorporation efficiency.

How do I submit my project?

Simply upload your gene sequence (FASTA format) or plasmid map through our online portal. Our scientific team will review your sequence for expression compatibility and provide a quote within 24 hours. For custom requirements or complex projects, you can schedule a consultation with our technical team.

What quality control is performed?

Every reaction includes thorough quality control. Standard QC includes SDS-PAGE analysis to verify expression and purity, concentration measurement by absorbance or BCA assay, and activity assays where applicable. Additional options include Western blot confirmation, mass spectrometry verification, and endotoxin testing for proteins intended for downstream biological applications.

Request a Quote

E. coli Cell-Free Protein Synthesis

Why Choose Us

Rapid Expression

Toxic Protein Friendly

Flexible Engineering

Why Choose Our E. coli CFPS Service

Rapid Expression

Toxic Protein Friendly

Flexible Engineering

Scalable Platform

Quality Guaranteed

Cost Effective

Ready to Accelerate Your Protein Research?

Our CFPS Technology

E. coli S30 Extract Platform

Optimized Reaction Buffer

Template Requirements

Quality Control

Purification Options

Service Specifications

Our Service Workflow

Template Submission

Reaction Setup

Expression

Quality Analysis

Delivery

Diverse Applications Across Biotechnology

Research Applications

High-Throughput Screening

Protein Engineering

What Our Clients Say

Scientific Foundation

Cell-Free PURE System: Evolution and Achievements

Adeno-Associated Virus 5 Protein Particles Produced by E. coli Cell-Free Protein Synthesis

Complex Dependence of Escherichia coli-based Cell-Free Expression on Sonication Energy During Lysis

Activation of Energy Metabolism through Growth Media Reformulation Enables a 24-Hour Workflow for Cell-Free Expression

Differentially Optimized Cell-Free Buffer Enables Robust Expression from Unprotected Linear DNA in Exonuclease-Deficient Extracts

Frequently Asked Questions

Ready to Start Your Project?

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