Blood Cell Stabilization Formulation for Ambient Storage
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Solution Overview
Problem
Current compositions and methods for stabilizing blood cells at ambient temperatures fail to maintain metabolic activity and morphology, making it difficult for subsequent analysis using automated cell separators and counters, and do not provide long-term stability without refrigeration or freezing.
Innovation Solution
Formulations comprising a pH buffer, a chelating agent, and a peptide, such as EDTA and di- or tri-peptides like Ala-Gln or Gly-Gly, that stabilize metabolically-active cells like leukocytes, erythrocytes, and circulating tumor cells, maintaining their native conformation and viability for extended periods at room temperature.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Temperature
If current compositions are used to stabilize blood cells at ambient temperatures, then storage without refrigeration is enabled, but metabolic activity and cell morphology are not maintained
Solution Approach 1:
The patent applies parameter changes by modifying the chemical composition parameters of the stabilization formulation. Specifically, it uses a pH buffer维持在7.2-7.4 to preserve enzymatic activity, adds 5-20 mM calcium ions to maintain cell membrane integrity and signaling functions, includes 1-10 mM magnesium ions for enzyme cofactor functions, and incorporates 0.1-1.0 mM EDTA to chelate excess metal ions that could cause oxidative damage. These coordinated parameter adjustments enable ambient temperature storage while maintaining metabolic activity.
Solution Approach 2:
The patent employs composite materials by creating a multi-component stabilization formulation that combines buffer substances (e.g., HEPES, MOPS), chelating agents (EDTA), metal ions (calcium, magnesium), and organic additives. This composite approach synergistically addresses multiple cellular requirements: pH buffering maintains protein structure, chelating agents prevent oxidative stress, and metal ions support enzymatic functions, collectively preserving metabolic activity at ambient temperatures.
2Temperature
If current compositions are used to stabilize blood cells, then ambient temperature storage is possible, but cell size and morphology are not retained
Solution Approach 1:
The patent maintains cell morphology by controlling physical chemical parameters in the formulation. The pH buffer (7.2-7.4) prevents protein denaturation and maintains cytoskeletal structure. Calcium ions (5-20 mM) stabilize cell membrane integrity and prevent cell lysis. The osmolarity is adjusted to isotonic conditions to prevent water influx or efflux that would alter cell size. These parameter controls preserve native cell shape and size for automated analysis compatibility.
3Temperature
If current compositions are used for cell stabilization, then ambient temperature storage is enabled, but stability is limited to short periods
Solution Approach 1:
The patent applies preliminary action by pre-loading the blood sample with protective agents before storage begins. The formulation is added immediately upon sample collection, establishing protective conditions in advance. Calcium and magnesium ions are pre-present to maintain membrane integrity and enzymatic function from the start. The pH buffer is pre-adjusted to prevent acidification that would occur during storage. This preliminary protection enables extended stability without refrigeration.
Solution Approach 2:
The patent converts potentially harmful ambient temperature conditions into beneficial effects through the formulation. The elevated temperature that would normally accelerate degradation is countered by the chelating agent EDTA that binds metal ions involved in oxidative damage. The buffer system converts potential pH drift from metabolic activity into a stable environment. Thus, ambient temperature storage becomes feasible by converting thermal energy that would cause damage into a controlled environment that preserves cells for extended periods.
4Reliability
If refrigeration or freezing is used for blood sample storage, then cell stability is maintained, but metabolic activity and native conformation are lost
Solution Approach 1:
The patent changes the temperature parameter from refrigerated (4°C) or frozen conditions to ambient temperature (20-25°C), which physiologically maintains enzyme activity and metabolic processes. Concurrently, it adjusts the chemical parameters by adding protective agents (buffers, chelators, metal ions) that compensate for the lack of cold stabilization. This parameter transformation allows the sample to remain metabolically active while achieving stability through chemical protection rather than thermal suppression.
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 formulations ensure that at least 80% of the cells remain metabolically-active for up to 18 days at ambient temperatures, allowing for stable storage and analysis without the need for refrigeration, facilitating diagnostic and therapeutic applications.
Implementation Method 1
The formulations comprise a pH buffer, a chelating agent, and a peptide
Implementation Method 2
The formulations comprise a pH buffer, a chelating agent, and a peptide
Implementation Method 3
the membrane-bound, cell surface proteins expressed on the substantially stored cells retain their native conformation in the blood sample
Data Source
Figure 1A
Figure 1B
Figure 2A~2B
AI summary
The present invention relates to the stabilization of one or more metabolically-active cell in a blood sample at ambient temperatures. In particular, formulations, compositions, articles of manufacture, kits and methods for substantially stable storage of one or more metabolically-active cell in a blood sample at ambient temperatures are provided.