High-Throughput Cell-Free Protein Expression (HT-CFPS) Service

The High-Throughput Cell-Free Protein Expression (HT-CFPS) Service leverages the speed, flexibility, and open nature of Cell-Free Protein Synthesis (CFPS) to accelerate protein discovery. This platform utilizes automated liquid handling systems and miniaturized reaction formats (96- or 384-well plates) to synthesize hundreds to thousands of target proteins simultaneously. By eliminating the time and variability associated with live cell culture, HT-CFPS drastically reduces project timelines for large-scale screening and prototyping, making it a critical tool for synthetic biology, directed evolution, and early-stage drug target validation .

CD Biosynsis offers highly flexible HT-CFPS solutions, compatible with various lysates including E. coli, WGE, and RRL, allowing clients to select the optimal system based on their protein's complexity and PTM requirements. Our automated pipeline ensures unparalleled precision, reproducibility, and rapid iteration , enabling the synthesis of diverse protein libraries, optimization of expression conditions, and direct functional screening of unpurified protein products.

Highlights

Core benefits of our HT-CFPS platform:

• Extreme Speed: Protein synthesis is complete in hours, enabling multiple rounds of screening and validation within a single day.
• Automation & Miniaturization: Utilizes robotic liquid handlers to dispense small reaction volumes, ensuring precision and minimizing reagent costs.
• Toxicity Tolerance: The open system successfully expresses proteins that are toxic or inhibitory to living cells, such as membrane proteins or regulatory kinases.
• Rapid Optimization: Ideal for quickly screening different buffer compositions, chaperone additions, or lysate types to identify optimal expression conditions for a target.

Applications

Key areas benefiting from high-throughput cell-free protein expression:

Directed Evolution & Enzyme Screening

Rapid synthesis and functional testing of large mutant enzyme libraries to identify variants with enhanced activity or stability.

Synthetic Biology Prototyping

Fast testing of genetic circuits, co-expression of multi-enzyme pathways, and optimization of stoichiometry in vitro .

Drug Target & Antibody Library Screening

Synthesis of multiple protein targets (e.g., scFvs, kinases) for immediate use in binding, functional, or activity assays.

Structural Biology Labeling (NMR)

Cost-effective, high-throughput incorporation of stable isotopes (15N, 13C) for structural analysis of protein libraries.

Key Features & Scalability

The comparative advantage of our automated HT-CFPS:

Robotic Precision

Automated liquid handling ensures reliable, reproducible dispensing across 96/384 wells, minimizing human error and variability.

Template Flexibility

Compatible with linear DNA (PCR product), plasmid DNA, or mRNA, enabling direct screening from gene synthesis or PCR amplification.

Lysate Agnosticism

We can implement E. coli (high yield), WGE (low background), or RRL (eukaryotic fidelity) based on the project's specific needs.

Assay Integration

Synthesis is often performed directly in assay-compatible microplates, allowing for immediate functional analysis without intermediate purification.

Custom Additives Screening

High-throughput testing of hundreds of different additives (e.g., detergents, lipids, cofactors, chaperones) for optimized folding.

Workflow

Our streamlined process for high-speed, parallel protein expression:

• Template Preparation: The supplied gene library is processed into the optimal template format (linear DNA or mRNA).
• Robotic Setup: Automated liquid handling dispenses lysate and template into 96- or 384-well plates with defined additives.
• In Vitro Expression: Plates are incubated (e.g., 2-12 hours at 25°C to 37°C) under optimized CFPS conditions.
• High-Throughput Assay: Synthesized proteins are directly quantified (e.g., fluorescence scan) and/or functionally assayed in the expression plate.
• Data Analysis: Comprehensive data package is delivered, including expression yields, activity heat maps, and sequence-to-function correlation analysis.

We provide essential assurance for high-quality HT-CFPS outcomes:

• Precision Guarantee: Low coefficient of variation (CV) across all wells, ensured by robotic precision.
• Functional Assay Integration: Expertise in linking expression directly to custom functional assays (e.g., FRET, Luminescence, SPR).
• Custom Buffer Screening: Guaranteed optimization of reaction conditions to maximize the yield or activity of difficult-to-express libraries.
• Scalability: Seamless transition of validated "hit" clones from small-scale screening to preparative-scale CFPS for purification.