Fibronectin Concentrate Standardization via Controlled Thawing
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Solution Overview
Problem
Current methods for preparing fibronectin concentrates from plasma are inefficient in standardizing fibronectin levels, which is crucial for promoting cell adhesion in cellular therapy and regenerative medicine products, as they often result in heterogeneity and require additional processing steps to achieve desired concentrations.
Innovation Solution
A method involving the separation of plasma from whole blood or apheresis, followed by freezing, thawing at controlled temperatures to precipitate high molecular weight proteins, filtering to remove fibrinogen, and irradiating the fibronectin-rich material, with subsequent concentration adjustment using cryoprecipitate AHF or plasma to achieve standardized fibronectin concentrations for use in creating a 3-dimensional matrix for cell cultures.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Manufacturing precision
If conventional methods are used to prepare fibronectin concentrates from plasma, then fibronectin can be obtained, but the fibronectin levels are heterogeneous and require additional processing steps to achieve desired concentrations
Solution Approach 1:
The patent applies parameter changes by controlling the thawing temperature parameter (refrigerated temperatures versus room temperature) to induce selective precipitation of high molecular weight proteins including fibronectin. This temperature parameter change creates a standardized concentration of fibronectin in the cryoprecipitate supernatant without requiring additional processing steps, directly resolving the contradiction between manufacturing precision and device complexity
Solution Approach 2:
The patent utilizes phase transitions by freezing plasma and then thawing it at controlled refrigerated temperatures to cause high molecular weight proteins to precipitate out of solution. This phase transition process naturally concentrates fibronectin in the supernatant, achieving standardized concentrations through the physical phase change rather than through complex additional processing steps
2Manufacturing precision
If plasma is frozen and thawed at refrigerated temperatures to precipitate high molecular weight proteins, then fibronectin concentrate is obtained, but the process requires extended time periods
Solution Approach 1:
The patent optimizes the thawing temperature parameter to refrigerated temperatures (specifically 2-8°C) which balances two competing factors: low enough to induce effective precipitation of high molecular weight proteins for standardized fibronectin concentration, but high enough to prevent complete solidification that would require extended thawing times. This parameter optimization resolves the contradiction between manufacturing precision and time loss by finding the optimal temperature window
Solution Approach 2:
The patent applies preliminary action by pre-freezing the plasma before thawing, which ensures that the plasma is in a controlled frozen state ready for standardized thawing processing. This preliminary freezing step allows for consistent and reproducible fibronectin concentration achievement during the subsequent controlled thawing period, reducing variability and potential need for reprocessing
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 ensures consistent fibronectin concentrations, enhancing cell adhesion and expansion in cultures by providing a standardized fibronectin-rich product, optimizing its use in cellular therapy and regenerative medicine applications.
Implementation Method 1
Plasma, whether derived from a WB unit or an apheresis procedure, is frozen and stored at temperatures at or below −20° C.
Implementation Method 2
When plasma is thawed at cold temperatures, the dissolved high molecular weight proteins and clotting factors become solids and fall (precipitate) out of the plasma solution
Implementation Method 3
Plasma is separated from whole blood (WB) units using a centrifugal force to create a division between the red blood cells (RBCs), white blood cells (WBCs), and plasma fractions of the unit
Implementation Method 4
Pressure is applied to the centrifuged WB bag to push plasma into an empty bag attached to the blood pack
Data Source
AI summary
Fibronectin enhances growth of adherent cells in culture. Methods herein involve producing human-derived fibronectin concentrate from the cryo-precipitation of plasma. Fibronectin concentrates from many donors are be pooled, filtered, and irradiated, followed by adjustment of the fibronectin concentration to fit individual customer needs. Aliquots of the final fibronectin Concentrate product can be used in cell therapy and regenerative medicine cell cultures.