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Søren Blirup-Jensen
Protein Standardization I: Protein Purification. Procedure for the Purification of Human Prealbumin, Orosomucoid and Transferrin as Primary Protein Preparations
A procedure for the purification of human prealbumin,
orosomucoid and transferrin as primary protein
preparations has been developed. The procedure describes
in detail the chemicals, the fractionation
equipment and the purification of the three proteins
from the same starting material (a 90-donor plasma
pool). The fractionation steps involve methods like
salting out, various anion and cation exchange chromatographies,
preparative electrophoresis, and finally,
size chromatography. Only mild and highly reproducible
fractionation methods are used in order to
obtain high recoveries. All of these provide the best
guarantee that no serious subfractionation has taken
place. At the same time, high recoveries are a necessity
for the pure protein to be representative of the
same protein
The pure proteins are produced as liquid calibrators (primary protein preparations) dissolved in one electrolyte (0.1 mol/l KCl) and stored in sealed glass ampoules at ?80 °C. These three pure proteins were used as primary reference preparations in the certification of the international reference preparation CRM 470.
Clinical Chemical Laboratory Medicine, Walter de Gruyter
Print ISSN: 1434-6621
Volume: 39, 11/2001
Pages: 1076 - 1089