Process Variations
The Gerlough method, developed in 1955 improved process economics by reducing the consumption of ethanol. Instead of 40% in certain steps, Gerlough used 20% ethanol for precipitation. This is especially used for Fractions II and III. In addition, Gerlough combined the two fractions with IV into one step to reduce the number of fractionations required. While this method provedless expensive, it was not adopted by industry because of this combination of fractions II, III, and IV, for fear of mixing and high impurities.
The Hink method developed in 1957. This method gave higher yields through recovery of some of the plasma proteins discarded in the Fractions of IV. The improved yields, however, balanced by the lower purities obtained, within the 85% range.
The Mulford method, akin to the Hink, used the fractions II and III supernatant as the last step before finishing and heat treatment. The method combined fractions IV and V, but in this case, the albumin would not be as pure, although the yields may be higher.
Another variation was developed by Kistler and Nitschmann, to provide a purer form of albumin, even though offset by lower yields. Similar to Gerlough, the Precipitate A, which is equivalent to Cohn’s Fraction II and III, was done at a lower ethanol concentration of 19%, but the pH, in this case, was also lower to 5.85. Also similar to Gerlough and Mulford, the fraction IV was combined and precipitated at conditions of 40% ethanol, pH of 5.85, and temperature of −8 degrees C. The albumin, which is recovered in fraction V, is recovered in Precipitate C at a pH adjustment to 4.8. Similar to the Cohn Process, the albumin is purified by extraction into water followed by precipitation of the impurities at 10% ethanol, pH 4.6, and −3 degrees C. Akin to the Cohn Process, the precipitate formed here is filtered out and discarded. Then Precipitate C (fraction V) is reprecipitated at pH 5.2 and stored as a paste at −40 degrees C. This process has been more widely accepted because it separates the fractions and makes each stage independent of each other.
| Precipitate: | A | B | C |
|---|---|---|---|
| Ethanol %: | 19 | 40 | 40 |
| pH: | 5.85 | 5.85 | 4.8 |
| Temperature (degs C) | −3 | −8 | −8 |
Another variation involved a heat ethanol fractionation. It was originally developed to inactivate the hepatitis virus. In this process, recovery of high yield, high purity albumin is the most important goal, while the other plasma proteins are neglected. In order to make sure the albumin does not denature in the heat, there are stabilizers such as sodium octanoate, which allow the albumin to tolerate higher temperatures for long periods. In heat ethanol, the plasma is heat treated at 68 degrees C with sodium octanoate with 9% ethanol at pH of 6.5. This results in improved albumin recovery with yields of 90%, and purities of 100%. It is not nearly as expensive as cold ethanol procedures such as the Cohn Process. One drawback is the presences of new antigens due to possible heat denaturation of the albumin. In addition, the other plasma proteins have practical uses and to neglect them would not be worth it. Finally, the expensive heat treatment vessels offset the lower cost compared to the cold ethanol format that do not need it. For these reasons, several companies haven not adopted this method even though it has the most impressive results. However, one prominent organization that uses it is the German Red Cross.
The latest variation was developed by Hao in 1979. This method is significantly simplified compared to the Cohn Process. Its goal is to create high albumin yields as long as albumin is the sole product. Through a two stage process, impurities are precipitated directly from fractions II and III supernatant at 42% ethanol, pH 5.8, temperature −5 degrees C, 1.2% protein, and 0.09 ionic strength. Fraction V is precipitated at pH 4.8. Fractions I, II, III, and IV are coprecipitated at 40% ethanol, with pH of 5.4 to 7.0, and temperature −3 to −7 degrees C. Fraction V is then precipitated at pH 4.8 and −10 degrees C. The high yields are due to a combination of a simplified process, with lower losses due to coprecipitation, and use of filtration. Higher purities were also achieved at 98% because of the higher ethanol levels, but the yields were lowered with the high purity.
More recent method involve the use of chromatograpy.
Read more about this topic: Cohn Process
Famous quotes containing the words process and/or variations:
“Thinking is seeing.... Every human science is based on deduction, which is a slow process of seeing by which we work up from the effect to the cause; or, in a wider sense, all poetry like every work of art proceeds from a swift vision of things.”
—Honoré De Balzac (1799–1850)
“I may be able to spot arrowheads on the desert but a refrigerator is a jungle in which I am easily lost. My wife, however, will unerringly point out that the cheese or the leftover roast is hiding right in front of my eyes. Hundreds of such experiences convince me that men and women often inhabit quite different visual worlds. These are differences which cannot be attributed to variations in visual acuity. Man and women simply have learned to use their eyes in very different ways.”
—Edward T. Hall (b. 1914)