UV/Vis Spectroscopy Quantitative Models for Bioprocess Concentration Control
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Solution Overview
Problem
Current methods for determining substance concentrations in bioprocessing, particularly protein and DNA, face challenges such as limited dynamic range, cross-sensitivity between DNA and protein, and saturation issues in UV/vis spectroscopy, which restrict accurate measurement across a wide concentration range and require frequent adjustment of optical path lengths.
Innovation Solution
The method employs multivariate data analysis to generate quantitative models based on absorption and intensity spectra from multiple concentration samples, allowing for the projection of undetermined concentrations using UV/vis spectroscopy with a single measurement cell of constant optical path length, enabling continuous monitoring and differentiation of substances with overlapping absorption bands.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Device complexity
If a single measurement cell with constant optical path length is used, then device complexity is reduced and ease of operation is improved, but measurement precision deteriorates due to saturation at high concentrations and limited dynamic range
Solution Approach 1:
The patent transitions from single-wavelength measurement to multi-wavelength spectral measurement, adding the wavelength dimension to the measurement space. By recording entire absorption spectra rather than single wavelength values, the system can differentiate substances with overlapping absorption bands and avoid saturation effects through selective wavelength analysis.
Solution Approach 2:
The patent changes the measurement parameter from single-wavelength absorbance to multi-wavelength spectral information. By analyzing the entire absorption spectrum and using multivariate data analysis, the system can determine concentrations across a wide dynamic range (0-200 g/L) without hardware adjustments, resolving the contradiction between simple measurement cell design and measurement precision.
2Measurement precision
If broadband spectroscopy with multivariate data analysis is used, then measurement precision is improved across wide concentration ranges, but device complexity increases due to need for sophisticated data processing and modeling
Solution Approach 1:
The patent performs preliminary action by generating quantitative models offline using multivariate data analysis on calibration data. These models are stored and then applied during actual measurements, separating the complex model generation phase from the simple measurement phase. This allows sophisticated analysis capabilities while keeping the operational system simple.
Solution Approach 2:
The patent creates mathematical models that copy the relationship between spectral data and concentration. Instead of performing complex physical measurements for each concentration determination, the system uses pre-established mathematical relationships (models) to predict concentrations from spectral data, simplifying the operational complexity.
3Measurement precision
If UV sensors with narrowband excitation are used, then measurement precision at specific wavelengths is improved, but adaptability deteriorates due to saturation at low concentrations and inability to measure wide concentration ranges
Solution Approach 1:
The patent makes the measurement system universal by using broadband light sources and detectors that can measure across the entire UV-Vis spectrum. This single system can handle all concentration ranges (0-200 g/L) and differentiate multiple substances (proteins, DNA, host cell proteins) without requiring different hardware configurations, achieving multi-functionality.
Solution Approach 2:
The patent adds the wavelength dimension to measurements, transitioning from single-wavelength narrowband detection to multi-wavelength broadband detection. This allows the system to avoid saturation by selecting appropriate wavelengths and to differentiate substances based on their unique spectral fingerprints, greatly enhancing adaptability across concentration ranges.
4Measurement precision
If optical path length is adjusted frequently to accommodate different concentration ranges, then measurement precision is maintained, but productivity decreases due to frequent hardware adjustments and interruptions
Solution Approach 1:
The patent performs preliminary action by establishing quantitative models that cover the entire expected concentration range (0-200 g/L) before actual process monitoring begins. During production, these pre-established models can be applied continuously without interruption, eliminating the need for frequent optical path adjustments and maintaining both precision and productivity.
Solution Approach 2:
The patent introduces dynamic wavelength selection based on concentration levels. The system automatically adapts which wavelengths to analyze based on the measured spectrum, allowing a single optical path length to effectively handle all concentration ranges. This dynamic software-based adaptation replaces static hardware adjustments, maintaining productivity.
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
This approach allows for precise and continuous monitoring of protein and DNA concentrations across a wide range, reducing cross-sensitivity and avoiding saturation, enabling efficient process control and quality assurance in bioprocessing without the need for hardware changes or frequent adjustments.
Implementation Method 1
spectrometers that are designed to record spectral information generally use broadband emitting light sources (also called broadband light sources)
Implementation Method 2
Concentrations of substances are typically determined or measured using optical spectroscopy, in particular using UV/vis absorption spectroscopy
Implementation Method 3
the molecules of the substance of a sample of the fermentation broth are generally excited with individual discrete wavelengths... and the absorbance or extinction, or alternatively the intensity, is determined
Data Source
AI summary
Method for open-loop or closed-loop control of a process, in particular a downstream bioprocess, based on the projection of an unknown concentration of at least one substance in a sample using spectroscopy, in particular UV/vis spectroscopy, comprising the steps: Detect spectrums of a plurality of concentration samples, wherein at least two concentration samples have differing concentrations of the substance; generate several quantitative models based on the spectrums of the concentration samples, wherein the models each have a mapping of at least one spectral measurand of the spectrums to concentrations in concentration ranges, wherein the concentration ranges of two models are not identical; detect at least one sample spectrum of the sample; map the sample spectrum to at least one quantitative model of the generated quantitative models; apply the at least one quantitative model that was mapped to the sample spectrum against the sample spectrum to determine a projected value for the unknown concentration; and apply open-loop and/or closed-loop control of the process for at least one parameter based on the projected value for the undetermined concentration.