Process Details
During the operations, the ethanol concentration change from zero initially to 40%. The pH decreases from neutral at 7 to more acidic at 4.8 over the course of the fractionation. The temperature starts at room temperature and decreases to −5 degrees Celsius. Initially, the blood is frozen. There are five major fractions. Each fraction ends with a specific precipitate. These precipitates are the separate fractions.
Fractions I, II, and III are precipitated out at earlier stages. The conditions of the earlier stages are 8% ethanol, pH 7.2, −3 °C, and 5.1% protein for Fraction I; 25% ethanol, pH of 6.9, −5 °C, and 3% protein. The albumin remains in the supernatant fraction during the solid/liquid extraction under these conditions. Fraction IV has several unwanted proteins that need to be removed. In order to do this, the conditions are varied in order to precipitate the proteins out. The conditions to precipitate these proteins are raising the ethanol concentration from 18 to 40% and raising the pH from 5.2 to 5.8. Finally, albumin is located in fraction V. The precipitation of albumin is done by reducing the pH to 4.8, which is near the pI of the protein, and maintaining the ethanol concentration to be 40%, with a protein concentration of 1%. Thus, only 1% of the original plasma remains in the fifth fraction.
However, albumin is lost at each process stage, with roughly 20% of the albumin lost through precipitation stages before fraction V. In order to purify the albumin, there is an extraction with water, and adjustment to 10% ethanol, pH of 4.5 at −3 °C. Any precipitate formed here is done so by filtration and is an impurity. These precipitates are discarded. Reprecipitation, or repetition of the precipitation step in order to improve purity, is done so by raising ethanol concentration back to 40% from the extraction stage. The pH is 5.2 and it is conducted at −5 °C. Several variations of Cohn fraction were created to account for lower cost and higher yield. Generally, if the yield is high, the purity is lowered, to roughly 85-90%.
| Fraction #: | Fraction I | Fraction II | Fraction III | Fraction IV | Fraction V |
|---|---|---|---|---|---|
| Ethanol %: | 8 | 25 | 18 | 40 | 40 |
| pH: | 7.2 | 6.9 | 5.2 | 5.8 | 4.8 |
| Temperature (°C) | −3 | −5 | −5 | −5 | −5 |
| Protein fraction (%): | 5.1 | 3 | 3 | 3 | 1 |
Read more about this topic: Cohn Process
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