Plasma Protein Separation via pH Gradient
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Solution Overview
Problem
Current plasma fractionation methods, such as the Cohn Cold Fractionation Process, are inefficient and can denature proteins, requiring extensive purification processes and the use of alcohol or other buffers, which complicates the separation of medically important proteins like Immunoglobulin G and AFOD from plasma fractions.
Innovation Solution
A pH gradient method is employed to separate proteins from plasma fractions III and IV using water for injection, without alcohol or buffers, involving pH adjustment, centrifugation, filtration, and low pH incubation to produce new formulations effective against Hepatitis C and HIV viruses.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Manufacturing precision
If the Cohn Cold Fractionation Process is used to separate proteins from plasma, then protein fractions can be obtained, but the process is complex and requires extensive purification steps including cryoprecipitation, nanofiltration, solvent detergent treatments, and incubation
Solution Approach 1:
The patent applies parameter changes by systematically varying pH levels and ethanol concentrations to achieve protein separation. Instead of using complex multi-step purification processes, the invention modifies solution parameters (pH from 2.0-3.0 to 6.0-7.0, ethanol concentration from 10% to 50%) to selectively precipitate and separate different protein fractions, thereby simplifying the overall process while maintaining separation efficiency
Solution Approach 2:
The invention extracts and removes specific protein fractions from plasma by selective precipitation. By adjusting pH and ethanol concentration, target proteins (such as immunoglobulins, albumin, and alpha-1 antitrypsin) are selectively precipitated and separated from the plasma mixture, allowing direct recovery of purified protein fractions without requiring extensive additional purification steps
2Manufacturing precision
If conventional fractionation methods are used, then proteins can be separated into fractions, but protein denaturation occurs requiring extensive purification processes
Solution Approach 1:
The patent carefully controls parameter changes to prevent protein denaturation. By gradually adjusting pH within specific ranges (2.0-3.0 to 6.0-7.0) and controlling ethanol concentration (10%-50%), the method maintains protein structural integrity while achieving separation. The controlled parameter changes ensure proteins remain in their native conformation throughout the fractionation process
Solution Approach 2:
The invention applies beforehand cushioning by maintaining optimal temperature conditions (4°C to 25°C) and using gentle, gradual parameter adjustments during the fractionation process. These pre-established protective conditions prevent protein denaturation before it can occur, ensuring protein stability is maintained throughout separation without requiring subsequent purification to remove denatured proteins
3Manufacturing precision
If alcohol or buffers are used in the fractionation process, then protein separation can be achieved, but the process becomes more complicated
Solution Approach 1:
The patent achieves protein fractionation by changing solution parameters (pH and ethanol concentration) rather than relying on complex buffer systems. By systematically varying pH from 2.0-3.0 to 6.0-7.0 and ethanol concentration from 10% to 50%, the method achieves effective protein separation using simple, controllable parameter adjustments, thereby simplifying the manufacturing process while maintaining fractionation precision
Solution Approach 2:
The invention discards complex buffer systems and replaces them with simpler ethanol-based precipitation methods. By recovering proteins through selective precipitation controlled by pH and ethanol concentration, the process eliminates the need for extensive buffer exchanges and complex purification steps, thereby simplifying manufacturing while maintaining effective protein fractionation
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 simplifies protein separation, prevents denaturation, and produces formulations that can stop and eradicate Hepatitis C and HIV-1/HIV-2 infections, while providing a more efficient and effective process for producing immunoglobulin and AFOD products.
Implementation Method 1
adjusting pH value to 1 and temperature from 1° C. to 30° C. to form a resultant suspension
Implementation Method 2
centrifuging the resultant suspension at 6,000 rpm at 2-8° C. for 20 min; collecting a resulted paste (P1) and supernatant 1 (S1)
Data Source
AI summary
The present subject matter is directed to a method for separating proteins of plasma using pH adjustment including the steps of reconstituting Fraction III, Fraction IV, or plasma paste, in water for injection; adjusting pH value to 1 and temperature from 1° C. to 30° C.; centrifuging the resulting suspension at 6,000 rpm at 2-8° C. for 20 min; collecting the resulting paste 1 (P1) and supernatant 1 (S1); reconstituting P1 in WFI and adjust the pH to 2; and repeating step 3) to step 5) until the pH of supernatant reaches 14. According to the method, a new formulation of immunoglobulin is prepared from plasma Fraction III and Fraction IV.