High-Throughput Cell-Free Protein Screening Service for Functional Assays and Hit Identification

The High-Throughput Cell-Free Protein Screening (HTP-CFPS) Service integrates the rapid synthesis capability of Cell-Free Protein Synthesis (CFPS) with miniaturized, automated functional analysis. This service moves beyond simple expression, focusing on fast, parallel screening of protein libraries directly in microplates (96- or 384-well) immediately following synthesis. This approach dramatically accelerates the identification of novel enzyme variants, binding partners, or drug targets based on specific functional criteria.

CD Biosynsis utilizes state-of-the-art robotic systems and optimized lysate platforms (E. coli, WGE, RRL) to manage vast libraries. Our HTP-CFPS platform allows for direct, in-well functional measurement—such as enzyme kinetics, binding affinity, or protein-protein interactions —from unpurified protein lysate. This eliminates time-consuming purification steps during the discovery phase, providing rapid, quantitative data on thousands of protein variants to drive directed evolution and drug discovery pipelines.

Highlights

Core benefits of our HTP-CFPS Screening Service:

• Direct Functional Screening: Assay proteins immediately after synthesis in the reaction well, bypassing purification for initial screening.
• Fast Hit Identification: Rapid synthesis and assay integration allow for screening of thousands of clones per day, accelerating the discovery cycle.
• Toxicity & Solubility Control: Screen toxic targets or optimize solubility conditions across many different buffer formulations simultaneously.
• Library Compatibility: Ideal platform for testing large, diverse libraries derived from directed evolution or genomic/cDNA sources.

Screening Applications

Critical applications where our HTP-CFPS platform delivers accelerated results:

Enzyme Kinetics and Activity Screening

High-throughput measurement of kcat and Km values across enzyme mutant libraries to identify enhanced biocatalysts.

Protein-Protein Interaction (PPI) Assays

Synthesizing interaction partners (e.g., baits and preys) in different combinations to map interaction networks or screen inhibitors.

Binding and Affinity Screening

Rapidly producing labeled receptors or ligands for high-throughput screening of small molecule or antibody candidates.

PTM Functionality and Processing

Screening the effect of mutations or cofactors on PTM status (e.g., phosphorylation or cleavage efficiency) and resulting activity.

Key Features & Sensitivity

Key technical advantages of our automated CFPS screening platform:

Integrated Detection

Supports fluorescence, luminescence, FRET/BRET, and colorimetric readouts directly in the expression well.

High Signal-to-Noise

Utilization of low-background lysates (like WGE or nuclease-treated RRL) ensures high sensitivity for detecting subtle activity changes.

Multiplexing Capability

Ability to screen multiple templates or cofactors in parallel using advanced plate layouts and robotic fluidics.

Automation Reliability

Robotic handling ensures consistent reaction setup and assay performance across thousands of data points.

Template-to-Hit Linkage

Direct linking of functional results back to the source gene sequence for immediate hit identification and re-synthesis.

Workflow

Our streamlined process for functional protein screening and hit identification:

• Template Library Preparation: Client-supplied library or synthesized gene library is converted to linear DNA or mRNA template.
• Robotic Expression Setup: Automated liquid handling dispenses template and lysate into microplates (96/384 format).
• In Vitro Synthesis: Incubation for 2-12 hours generates the protein library in the reaction wells.
• Functional Assay Integration: Specific substrate, binding partner, or fluorescent probe is added, and the functional signal is measured by a plate reader.
• Hit Identification & Validation: Results are analyzed, hits are ranked by activity/binding affinity, and the corresponding gene templates are identified for validation.

We provide essential assurance for high-quality screening outcomes:

• Quantitative Data: Delivery of quantitative functional data (e.g., RFU, RLU, or Kcat) for every screened variant.
• Low CV Guarantee: Guaranteed low coefficient of variation across replicate wells for high data confidence.
• Custom Assay Development: Expertise in adapting complex functional assays for compatibility with the cell-free environment.
• Sequence Traceability: Guaranteed linkage of functional results back to the original gene sequence for seamless follow-up.