Variable Pathlength Fluorescence Protein Concentration Measurement
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Existing methods for measuring protein concentrations, especially in commercial production settings, face challenges such as contamination risks, unreliable results at high and low concentrations, and disruption to production workflows due to the need for sampling and sample preparation, which are exacerbated by the use of additives like dyes in colorimetric methods and the non-specificity of refractive index measurements.
Innovation Solution
A method and device utilizing a sample container with a transparent wall segment and focused excitation radiation that induces fluorescence, allowing for non-invasive measurement of protein concentrations without contamination, capable of varying pathlengths to accommodate a wide concentration range, using a monochromatic light source between 290-330 nm and a detector for induced radiation, which can be positioned on the same side as the excitation source to capture fluorescence directly.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If UV absorbance measurement at 280 nm is used with fixed pathlength, then measurement is simple and fast, but measurement precision deteriorates at very high and very low concentrations
Solution Approach 1:
The patent applies dynamics by making the pathlength variable rather than fixed. The measurement system dynamically adjusts the pathlength of light through the sample based on the concentration being measured, allowing accurate measurements across a wide concentration range from very low to very high concentrations without requiring manual intervention or dilution.
Solution Approach 2:
The patent changes the physical parameter of pathlength to resolve the measurement precision issue. By varying the pathlength parameter according to the sample concentration, the system maintains optimal measurement conditions across different concentration levels, preventing both the saturation effect at high concentrations and the detection limit issue at low concentrations.
2Measurement precision
If colorimetric methods with dyes are used, then measurement sensitivity is improved, but contamination risk and workflow disruption increase due to reagent addition
Solution Approach 1:
The patent extracts and eliminates the need for external reagents and dyes from the measurement process. By using intrinsic UV absorbance properties of proteins at 280 nm, the method removes harmful reagents entirely, preventing contamination risks and workflow disruptions associated with adding chemicals to the sample.
Solution Approach 2:
The protein sample serves itself in the measurement process by utilizing its own intrinsic UV-absorbing aromatic amino acids (tryptophan, tyrosine, phenylalanine) at 280 nm. This self-service approach eliminates the need for external reagents that could cause contamination, while maintaining measurement sensitivity through the natural optical properties of the protein.
3Ease of operation
If refractive index measurement is used, then measurement is non-specific to proteins, but measurement simplicity is improved
Solution Approach 1:
The patent utilizes optical absorbance changes at a specific wavelength (280 nm) rather than refractive index changes. This wavelength-specific absorbance method provides both simplicity and specificity, as proteins naturally absorb UV light at 280 nm due to their aromatic amino acid content, allowing straightforward measurement that is inherently specific to proteins without complex instrumentation.
4Measurement precision
If sample dilution is performed to extend measurement range, then measurement precision at high concentrations is improved, but device complexity and time consumption increase
Solution Approach 1:
The patent applies dynamics by implementing variable pathlength capability that automatically adapts to different concentration levels. This dynamic adjustment eliminates the need for manual dilution steps, reducing both device complexity and time consumption while maintaining measurement precision across the full concentration range.
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 accurate measurement of protein concentrations across a broad range, from very low to very high concentrations, without disrupting the production process, reducing contamination risks and improving measurement reliability and speed, while being suitable for continuous production settings.
Implementation Method 1
capturing a measurement value by measuring an induced radiation being induced by an interaction of the excitation radiation with the sample, wherein the interaction comprises fluorescence
Data Source
Figure 1
Figure 2
Figure 3~4
AI summary
A method for measuring a concentration value of a substance within a sample is provided. The method comprises the following steps: providing a sample container (4) with a transparent wall segment (10) on at least one side (S) of the sample container (4), providing a sample (2) having an unknown concentration of the substance in the sample container (4), providing an excitation radiation (R1) having a focal point (F) such that the excitation radiation reaches into the sample container (4) via the wall segment (10) such that the focal point (F) is located at a first distance (δ) from the wall segment (10), capturing a measurement value by measuring an induced radiation (R2) being induced by an interaction of the excitation radiation (R1) with the sample (2), wherein the interaction comprises fluorescence and wherein the induced radiation leaves the sample container (4) via said wall segment (10), and comparing the measurement value to a comparative value to derive a value indicative of the unknown concentration value.