UV Protein Concentration Imaging With Automated Multi-Wavelength Calibration
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Solution Overview
Problem
Current instruments for measuring protein concentrations in pharmaceuticals are limited by the Beer-Lambert Law's linearity, leading to errors, false positives, and false negatives due to reliance on fixed or variable path lengths, and are prone to mechanical issues and high costs.
Innovation Solution
A UV-based imaging system using automated multi-wavelength calibration to determine protein concentrations by projecting a multi-wavelength light beam, detecting it with a monochromator, and applying a multi-wavelength calibration model to calculate protein concentrations accurately over a wide range without mechanical errors.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Ease of operation
If fixed path length UV spectrometers are used to measure protein concentration, then the measurement process is simple, but the measurement precision deteriorates when absorbance is out of the linear range due to Beer-Lambert Law limitations
Solution Approach 1:
The patent transitions from single-wavelength absorbance measurement to multi-wavelength spectral measurement. By measuring absorbance across multiple wavelengths (200-400nm) rather than a single wavelength, the system creates a spectral fingerprint that enables accurate concentration determination even when individual wavelengths fall outside the linear range of Beer-Lambert Law.
Solution Approach 2:
The patent changes the measurement parameter from single absorbance value at fixed wavelength to multi-wavelength absorbance spectrum. The system measures absorbance at multiple wavelengths simultaneously and uses the spectral pattern (ratios, shapes) rather than absolute absorbance values, allowing accurate measurement across a wider concentration range without requiring samples to fall within the linear absorbance range.
2Measurement precision
If serial dilutions are performed to bring sample concentration into dynamic range, then measurement precision may improve, but the process becomes time consuming and error prone
Solution Approach 1:
The system automatically determines optimal wavelengths and performs calibration without manual intervention. The automated workflow selects calibration wavelengths, acquires spectral data, fits calibration curves, and calculates concentrations without requiring manual sample dilution or preparation, eliminating time-consuming manual steps while maintaining accuracy.
Solution Approach 2:
The system performs multi-wavelength spectral scanning and automated calibration curve fitting in advance to establish the measurement model. By pre-determining the spectral characteristics and calibration parameters across the full concentration range, the system eliminates the need for time-consuming serial dilutions during actual sample measurement.
3Measurement precision
If variable path length instruments are used to measure high protein concentration directly, then sample dilution is eliminated, but the device complexity and cost increase due to mechanical movement requirements
Solution Approach 1:
The patent replaces the mechanical variable path length system with a fixed path length cell and multi-wavelength optical system. Instead of mechanically adjusting the light path to accommodate different concentrations, the system uses spectral measurement at multiple wavelengths with a fixed cell, eliminating mechanical complexity while maintaining the ability to measure across wide concentration ranges through automated multi-wavelength calibration.
4Adaptability or versatility
If variable path length instruments are used, then direct measurement of high concentration is enabled, but measurement precision deteriorates due to mechanical errors and Fibrette manipulation
Solution Approach 1:
The patent eliminates mechanical path length adjustment mechanisms and Fibrette manipulation by using a fixed path length cell combined with multi-wavelength spectral measurement. The system achieves concentration range adaptability through optical (multi-wavelength) rather than mechanical means, thereby eliminating mechanical errors and improving measurement repeatability while maintaining the ability to measure across wide concentration ranges.
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
The system provides accurate protein concentration measurements over a wide range (0.1-126 mg/mL) with enhanced precision and reduces human error, eliminating the need for sample dilution and mechanical calibration, and is compatible with existing UV spectrometers.
Implementation Method 1
The absorbance value (A) is generally measured as a response factor to the light beam passing through a sample
Data Source
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AI summary
Ultraviolet (UV) based imaging method for determining protein concentrations of unknown protein samples based on automated multi-wavelength calibration. In various embodiments, a processor receives each of a standard set of wavelength data and an unknown set of wavelength data as recorded by a detector. Each standard set of wavelength data and unknown set of wavelength data defines a series of absorbance-to-wavelength value pairs across a first range of wavelengths selected from a range of a single-wavelength light beams of a UV spectra. The processor generates a multi-wavelength calibration model based on each of a first series of first absorbance- to-wavelength value pairs of the standard set of wavelength data. The processor implements the multi-wavelength calibration model to determine, for each unknown protein sample of the given unknown protein samples, a plurality of protein concentration values.