Vitamin B6 Status Detection via Erythrocyte PLP-DE Probe
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Solution Overview
Problem
Current methods for determining vitamin B6 status in subjects are limited by the instability of pyridoxal phosphate (PLP) in plasma samples, variability due to factors like inflammation and alcohol consumption, and the time-consuming processing required for functional assays.
Innovation Solution
An in vitro method involving the use of a probe with functionalized cofactors of PLP-dependent enzymes (PLP-DEs) to capture and label PLP-DEs in erythrocytes, followed by detection with anti-PLP-DE antibodies, allowing for rapid determination of vitamin B6 status.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If plasma PLP levels are measured using HPLC, then the vitamin B6 status can be determined, but the measurement precision is affected by PLP instability and binding to hemoglobin
Solution Approach 1:
The invention extracts and measures erythrocyte PLP levels separately from plasma components. By isolating the intracellular PLP measurement from plasma matrix effects and hemoglobin binding interference, the method achieves more reliable and precise vitamin B6 status assessment without the confounding factors present in plasma measurements.
2Measurement precision
If functional B6 biomarker tests are performed on fresh blood, then the vitamin status assessment is more accurate, but the processing time increases significantly
Solution Approach 1:
The invention performs preliminary isolation of erythrocytes from whole blood before functional analysis. By pre-separating the red blood cells containing the PLP-dependent enzymes from plasma and other components, the method enables subsequent rapid functional testing without requiring extensive fresh blood processing, thus reducing overall analysis time while maintaining accuracy.
3Adaptability or versatility
If multiple PLP-dependent enzymes are analyzed, then personalized vitamin B6 status parameters can be obtained, but the device complexity increases
Solution Approach 1:
The invention employs a universal functional assay approach where a single test system can analyze multiple PLP-dependent enzymes simultaneously. By using a common methodology to assess various enzymes (such as transaminases, decarboxylases, and other PLP-DEs), the system provides personalized vitamin B6 status parameters across different enzyme activities without requiring separate complex diagnostic platforms for each enzyme.
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 method provides a rapid, reliable, and personalized assessment of vitamin B6 status, independent of free PLP stability and fluctuations, and can be performed with minimal equipment, making it suitable for point-of-care testing.
Implementation Method 1
treating the lysate with a probe comprising at least one functionalized cofactor of PLP-dependent enzymes (PLP-DEs)
Implementation Method 2
capturing the labeled PLP-DEs with anti-PLP-DE-antibodies immobilized on a surface
Data Source
AI summary
The present invention relates to in vitro methods for determining the vitamin status of a subject, in particular health related vitamins, such as B2, B6, B12 and/or D3. The present invention further relates to a kit and an immunographic device for in vitro determining the vitamin status of a subject.