High-throughput label-free coherent anti-stokes raman scattering (CARS) platform for protein variant screening and characterization

Overview

This high-throughput Coherent Anti-Stokes Raman Scattering (CARS) spectroscopic platform accelerates the characterization, quality control, and ranking of recombinant protein variants. CARS generates chemically specific vibrational fingerprints that report secondary structure, folding status, aggregation, solubility, and local microenvironment—all without the need for fluorescent tags or antibodies. This label-free approach eliminates potential interference from labeling reagents, preserves native protein behavior, and streamlines the screening workflow.

The platform is designed for rapid parallel screening of many variants in multi-well or microfluidic formats, significantly reducing cycle time while supporting data-driven ranking and iterative optimization. It complements computational design and expression tools by delivering fast, quantitative experimental readouts of protein behavior under relevant conditions. Validation is performed on representative herbicide-target variant libraries, delivering clear metrics on throughput, time savings, and correlation with functional performance.

Technical specifications

Key features:

• Label-free CARS spectroscopy delivers chemically specific vibrational fingerprints of protein structure and state
• Reports secondary structure, folding status, aggregation, solubility, and local microenvironment in a single measurement
• Compatible with microbial, cell-free, and protoplast expression systems
• High-throughput parallel screening in multi-well or microfluidic formats
• Benchmarking against conventional solubility, stability, and activity assays to quantify correlation, throughput, and time savings
• Data pipeline converts spectral features into quantitative ranking scores for iterative variant prioritization

A 6-month feasibility study framework is in place to configure instrumentation and protocols for partner-specific proteins, establish spectral baselines for folded, aggregated, and unfolded states, perform parallel multi-well screening on defined variant libraries, and validate the approach against existing assay methods.

Technology readiness level

The platform is currently at the validation stage. A 6-month feasibility study has been designed to adapt and validate the high-throughput CARS platform for recombinant protein variants. The study includes configuring instrumentation and protocols, establishing spectral baselines for folded, aggregated, and unfolded states, performing parallel multi-well CARS screening, and benchmarking results against existing solubility, stability, and activity assays. The final deliverable is a complete technical report with performance metrics, recommended protocols, and clear recommendations for scaling if results are successful.