Preparative displacement electrophoresis (isotachophoresis) of proteins on cellulose columns.
This paper describes the separation of proteins by displacement electrophoresis on columns packed with cellulose powder as a stabilizing medium. Cellulose has virtually no molecular sieving properties and thus differs from dextran, polyacrylamide, and agarose in this respect. Therefore, without the risk of unstacking, columns packed with cellulose permit conventional elution of the protein zones and the use of a counter flow (to increase the effective length of the bed). For the same reason, electroosmotic flow is less disturbing. A continuous elution-migration technique adapted to suit the special requirements of displacement electrophoresis gave better separation than was obtainable by conventional elution. Normal human serum and a fresh hemolysate from human erythrocytes were used as samples. An expression for the volume velocity of the boundaries is derived. This parameter can be used to determine the maximum duration of a run and a suitable pump speed when continuous elution or a counter flow is employed. The special advantages of displacement electrophoresis in cellulose beds are discussed as well as general disadvantages of the displacement technique, including the risk that proteins precipitate during a run.
Institute of Biochemistry, University of Uppsala, Sweden.
G
G
Johansson
L G
LG
Ofverstedt
S
S
Hjertén
D001797:Blood Protein Electrophoresis; D002482:Cellulose; D004586:Electrophoresis / Q000295:instrumentation; D004591:Electrophoresis, Polyacrylamide Gel; D004912:Erythrocytes / Q000032:analysis; D011506:Proteins / Q000032:analysis*
D011506:Proteins; D002482:Cellulose