Biological Kit for Separating Electronegative LDL
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Solution Overview
Problem
Current methods for separating electronegative low-density lipoproteins (LDL-) from plasma are time-consuming and result in undesirable quality due to the use of density gradient media like iodixanol, which cannot be efficiently removed, rendering further analysis impossible.
Innovation Solution
A biological kit containing glycerol and specific reagents without organo-iodine additives is used for the separation of electronegative low-density lipoproteins, employing a one-step centrifugation method that reduces operation time and improves separation quality.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If conventional multi-step separation methods are used, then separation completeness is improved, but operation time increases to at least 96 hours
Solution Approach 1:
The separation process is divided into distinct functional steps: (1) pre-centrifugation to remove chylomicrons and other lipoproteins, (2) density gradient centrifugation to separate LDL from HDL, and (3) dialysis to remove iodixanol. This segmentation allows each step to be optimized independently, achieving complete separation in a streamlined workflow that reduces total operation time while maintaining reliability.
Solution Approach 2:
The method performs preliminary removal of chylomicrons and other interfering lipoproteins through pre-centrifugation before the main density gradient separation. This preliminary action clears the sample of interfering components, allowing the subsequent density gradient centrifugation to focus solely on separating LDL from HDL, thereby reducing the time required for the main separation process while ensuring complete separation.
2Loss of time
If density gradient medium like iodixanol is used for rapid separation, then operation time is reduced, but separation quality deteriorates due to iodixanol contamination
Solution Approach 1:
The method extracts and removes iodixanol from the separated lipoprotein portions through dialysis against buffer solutions. This extraction step selectively removes the iodixanol contaminant while preserving the integrity of the separated LDL and HDL, thereby maintaining high separation quality even when using rapid density gradient centrifugation with iodixanol medium.
Solution Approach 2:
The patent introduces dialysis as an intermediary step between the density gradient centrifugation and final sample analysis. This intermediary process serves as a mediator that removes iodixanol contamination without affecting the separated lipoproteins, enabling the use of rapid iodixanol-based separation while ensuring high quality results through the mediating dialysis step.
3Reliability
If multiple sequential separation steps are performed, then separation completeness is improved, but device complexity and process difficulty increase
Solution Approach 1:
The method merges the separation of different lipoprotein classes into a single density gradient centrifugation step that simultaneously separates LDL from HDL, rather than requiring multiple sequential separations. This merging is achieved by optimizing the density gradient conditions to create distinct density zones that allow concurrent separation of multiple lipoprotein types, reducing process complexity while maintaining complete separation through the unified approach.
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 method significantly reduces operation time and achieves high-quality separation of electronegative LDL, allowing for further analysis and identification of metabolism-related disorders without the limitations of iodixanol contamination.
Implementation Method 1
the current approaches to separate the electronegative low-density lipoproteins from the plasma
Implementation Method 2
density gradient medium, such as iodixanol
Data Source
AI summary
A biological kit for separating electronegative low density lipoproteins is disclosed. The biological kit includes a first reagent, a second reagent, a third reagent and a fourth reagent. The first reagent includes water. The second reagent includes a first buffer, a second buffer and a salt. The third reagent includes the first buffer, the second buffer, an organic compound and the salt. The fourth reagent includes the first buffer and the second buffer.