Supercritical Carbon Dioxide Cell Lysis for Platelet Extraction
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Solution Overview
Problem
Current methods for releasing intramembrane therapeutic components from mammalian cells, such as platelets, are inefficient, with sub-optimal lysis efficiency, degradation of proteins, and require multiple cycles or lengthy processes, failing to achieve high concentrations of bioactive components within 30 minutes without toxins.
Innovation Solution
Exposing mammalian cells to supercritical carbon dioxide (SCCO2) at specific pressures (1071 to 5000 PSI) and temperatures (31.1 to 45°C) for 1 to 30 minutes, which gently dissociates cellular membranes to release therapeutic components while preserving their integrity and bioactivity, followed by controlled off-gassing to prevent freezing.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If conventional lysis methods (freeze-thaw, ultrasonification, hypotonic solutions) are used to release intramembrane components, then some therapeutic components are released, but lysis efficiency remains sub-optimal (less than 50%) and proteins degrade or unfold
Solution Approach 1:
The patent applies parameter changes by transitioning carbon dioxide to a supercritical state (temperature above 31.1°C and pressure above 73 atm) to create a unique solvent environment that enables efficient membrane disassociation while preserving protein structure. This phase change allows the system to achieve both high lysis efficiency and maintain protein integrity simultaneously
Solution Approach 2:
Supercritical carbon dioxide serves as an intermediary substance that mediates the lysis process. It acts as a gentle solvent that can penetrate and disassociate cellular membranes without directly damaging therapeutic proteins, thereby resolving the contradiction between efficient component release and protein preservation
2Productivity
If multiple cycles of freeze-thaw or ultrasonic pulses are applied to increase lysis, then more components are released over time, but the process takes at least 30 minutes and may cause protein degradation
Solution Approach 1:
The supercritical fluid extraction process operates continuously, allowing efficient component release within a single 30-minute exposure window. The supercritical carbon dioxide maintains constant extraction pressure and temperature throughout the process, eliminating the need for repeated cycles and reducing total processing time while achieving high component release concentrations
Solution Approach 2:
The patent utilizes phase transitions of carbon dioxide (between supercritical and gaseous states) to control the extraction process. By adjusting pressure and temperature, the system can rapidly transition between extraction and release phases, achieving efficient component release in under 30 minutes without requiring multiple repetitive cycles
3Loss of time
If aggressive lysis methods are used to achieve rapid component release, then processing time is reduced, but toxins and cellular debris are released along with therapeutic components
Solution Approach 1:
By precisely controlling temperature and pressure parameters within specific ranges (temperature above 31.1°C, pressure above 73 atm), the supercritical carbon dioxide achieves effective membrane disassociation while maintaining a gentle extraction environment that does not release toxins or excessive cellular debris, thus resolving the contradiction between rapid processing and product purity
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 achieves at least 50% release of intramembrane components from platelets within 10 minutes, maintaining their therapeutic efficacy and bioactivity, significantly improving upon existing techniques by providing a residue-free product in less than 30 minutes.
Implementation Method 1
The cells are exposed to supercritical carbon dioxide (SCCO2) for 1 to 30 minutes, where the SCCO2 is maintained at a pressure of 1071 to 5000 pounds per square inch (PSI) and a temperature of 31.1 to 45 degrees Celsius during the exposure. The exposure disassociates the cellular membranes of the cells to release intramembrane components
Implementation Method 2
the SCCO2 is off-gassed where the SCCO2 is maintained above freezing temperatures, transiting directly to liquid and/or gaseous phase carbon dioxide without the formation of a solid (i.e. dry ice). This serves to protect the intramembrane components from freezing to preserve the integrity, conformation, and bioactivity of the intramembrane components
Data Source
AI summary
A method of producing the constituents of a therapeutic product from mammalian cells is described herein. Cells are isolated from a mammalian source. The cells are exposed to supercritical carbon dioxide (SCCO2) for 1 to 30 minutes, where the SCCO2 is maintained at a pressure of 1071 pounds per square inch (PSI) and a temperature of 31.1 to 45 degrees Celsius during the exposure. The exposure dissociates the cellular membranes of the cells to release intramembrane components therein to produce constituents of the therapeutic product. The mammalian cells may include at least one of platelets, stem cells, germ cells, and somatic cells. The methods described herein are particularly advantageous for releasing and capturing therapeutic intramembrane components from platelets and alpha-granules.