QCL Hyperspectral Imaging for Protein Deamidation Detection
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Solution Overview
Problem
Current methods for evaluating deamidation in proteins are limited in throughput and require complex, time-consuming processes, making it difficult to assess the stability and potential immunogenicity of protein therapeutics effectively.
Innovation Solution
A system and method using quantum cascade laser (QCL) microscopy with hyperspectral imaging (HSI) for real-time, high-throughput analysis of protein samples, allowing for the determination of deamidation, aggregation, and stability without separation techniques, and employing 2D IR correlation spectroscopy to analyze spectral data for predictive insights.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If current techniques such as HPLC, NMR and MS are used to evaluate deamidation, then measurement precision is improved, but productivity deteriorates due to low throughput and time-consuming processes
Solution Approach 1:
The patent replaces complex mechanical separation systems (HPLC, NMR, MS) with a simplified optical detection system using surface-enhanced Raman spectroscopy (SERS). This substitution maintains measurement precision for deamidation detection while dramatically improving throughput by eliminating time-consuming separation and fragmentation steps.
Solution Approach 2:
The patent extracts and isolates the specific spectral signatures of deamidated residues using SERS, separating the detection function from the complex analysis workflows of traditional methods. This extraction allows direct detection without requiring full protein separation or peptide mapping, thereby improving productivity.
2Measurement precision
If HPLC separation techniques are used to detect deamidation, then measurement precision is improved, but device complexity and loss of time increase due to complex and time-consuming processes
Solution Approach 1:
The patent replaces complex mechanical separation devices (HPLC systems with ion exchange or reverse phase columns) with a simple optical detection platform using SERS. This substitution maintains the ability to detect deamidation with high precision while eliminating the need for complex separation equipment and procedures.
Solution Approach 2:
The patent changes the detection parameter from separation-based physical properties to vibration-based spectral signatures. By monitoring characteristic Raman shifts of deamidated versus non-deamidated residues, the system achieves precise detection without requiring complex separation devices.
3Measurement precision
If MS technique with fragmentation and peptide mapping is used, then measurement precision is improved for isoaspartate detection, but productivity deteriorates and device complexity increases
Solution Approach 1:
The patent extracts the detection function directly from the intact protein using SERS, eliminating the need to fragment the protein into peptides as required by MS. This extraction approach maintains precision in detecting isoaspartate while dramatically improving speed by analyzing the full-length protein in a single step.
Solution Approach 2:
The patent replaces the complex MS fragmentation and mapping system with a direct optical detection method using SERS. This substitution allows precise isoaspartate detection through characteristic spectral changes without requiring protein fragmentation, thereby improving productivity and reducing device complexity.
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
Enables fast and accurate assessment of deamidation and aggregation in proteins, providing predictive insights into immunogenicity and stability, facilitating early selection of stable therapeutic candidates and reducing the risk of candidate withdrawal.
Implementation Method 1
obtain and analyze spectral data for proteins, including infrared (IR) spectra, such as IR spectra obtained using a quantum cascade laser ('QCL') microscope
Implementation Method 2
The system and methods provide real-time high-throughput hyperspectral imaging ('HSI') that allows for the monitoring of an array of proteins in solution during thermal stress
Implementation Method 3
The system uses a QCL transmission microscope with linear response detection based on first principle, accurate thermal control, and unique heated cell holder with a multiplexed array slide cell
Data Source
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AI summary
Characteristics of proteins, peptides, and/or peptoids can be determined via two-dimensional correlation spectroscopy and/or two-dimensional co-distribution spectroscopies. Spectral data of the proteins, peptides, and/or peptoids can be obtained with respect to an applied stress, such as thermal stress. Two-dimensional correlation spectroscopy can be used to generate two-dimensional synchronous and asynchronous plots. The asynchronous plot provides enhanced resolution and the sequential order of molecular events that occur as a function of the applied stress. Peaks may be identified in the asynchronous plot, and correlation of peaks that exhibit out-of-phase intensity changes can be used to determine the existence and extent of deamidation events.