Single Assay Cholesterol Sub-Fraction Measurement via Kinetic Enzyme Differentiation
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Solution Overview
Problem
Current methods for measuring cholesterol sub-fractions in blood plasma are inaccurate and require multiple assays, leading to increased costs and errors due to the need for physical separation techniques like centrifugation and electrophoresis.
Innovation Solution
A single assay method using a combination of lipase, oxidase, and peroxidase enzymes that convert cholesterol-containing lipoproteins to measurable products at different rates, allowing for the kinetic differentiation and measurement of HDL-C, LDL-C, and VLDL-C without precipitation or separation steps.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If physical separation techniques like centrifugation and electrophoresis are used to measure cholesterol sub-fractions, then measurement precision can be improved, but device complexity and loss of time increase
Solution Approach 1:
The patent extracts the measurement of different cholesterol sub-fractions (HDL-C, LDL-C, VLDL-C) from the complex physical separation process by using selective enzyme inhibition. Specific enzymes are inhibited by particular substances to measure each sub-fraction independently in a single assay mixture, eliminating the need for centrifugation and electrophoresis equipment.
Solution Approach 2:
The patent creates a universal assay system where a single reaction mixture can measure multiple cholesterol sub-fractions simultaneously. The same basic enzymatic reaction system, combined with selective inhibitors, serves as a multi-functional platform for determining HDL-C, LDL-C, and VLDL-C without requiring separate measurement systems or separation apparatus.
2Measurement precision
If multiple assays are performed to measure cholesterol sub-fractions, then measurement precision improves, but loss of time and productivity decrease
Solution Approach 1:
The patent merges the measurement of multiple cholesterol sub-fractions into a single assay procedure. By combining multiple enzymatic reactions in one reaction mixture and using selective inhibition strategies, the system determines HDL-C, LDL-C, and VLDL-C simultaneously, eliminating the need for sequential separate assays and reducing total measurement time.
Solution Approach 2:
The patent performs preliminary enzymatic reactions to convert different cholesterol sub-fractions into measurable forms before the actual measurement step. Enzymes such as lipoprotein lipase and hepatic triglyceride lipase are used in advance to hydrolyze triglycerides and prepare cholesterol esters for subsequent oxidative reactions, enabling rapid simultaneous determination of multiple sub-fractions.
3Measurement precision
If multiple assays are performed to measure cholesterol sub-fractions, then measurement precision improves, but device complexity and costs increase
Solution Approach 1:
The patent merges the measurement of multiple cholesterol sub-fractions into a single assay procedure. By combining multiple enzymatic reactions in one reaction mixture and using selective inhibition strategies, the system determines HDL-C, LDL-C, and VLDL-C simultaneously, eliminating the need for sequential separate assays and reducing total measurement time.
4Measurement precision
If physical separation techniques are used, then measurement precision improves, but loss of substance increases
Solution Approach 1:
The patent creates a universal assay system where a single reaction mixture can measure multiple cholesterol sub-fractions simultaneously. The same basic enzymatic reaction system, combined with selective inhibitors, serves as a multi-functional platform for determining HDL-C, LDL-C, and VLDL-C without requiring separate measurement systems or separation apparatus.
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 provides accurate and precise measurements of cholesterol sub-fractions in a single assay, reducing errors and costs by eliminating the need for multiple assays and physical separation techniques.
Implementation Method 1
A single assay method using a combination of lipase, oxidase, and peroxidase enzymes that convert cholesterol-containing lipoproteins to measurable products at different rates
Implementation Method 2
A single assay method using a combination of lipase, oxidase, and peroxidase enzymes that convert cholesterol-containing lipoproteins to measurable products
Implementation Method 3
convert cholesterol-containing lipoproteins to measurable products
Data Source
AI summary
Reagents, assays, methods, kits, devices, and systems for rapid measurement of cholesterol and cholesterol sub-fractions from a blood sample are provided. Total cholesterol, low density lipoprotein cholesterol, and high density lipoprotein cholesterol can be measured in a single assay using kinetic measurements, under conditions in which cholesterol sub-species are converted to a detectable product at distinct rates. The detectable product is measured at different times after assay initiation. A lipase, cholesterol esterase, cholesterol oxidase and a peroxidase may be used together to produce colored product in amounts directly proportional to the quantity of cholesterol converted. Methods for calculating very-low density lipoprotein cholesterol levels by further including triglyceride measurements are disclosed. Assays may be performed in a single reaction mixture, allowing more accurate and precise cholesterol determinations, including ratios of cholesterol sub-fractions to total cholesterol, at less expense, than would be expected by performing several different assays in different reaction mixtures.