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 molecularsieving properties and thus differs from dextran, polyacrylamide, and agarose in this respect. Therefore, without the r...

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Veröffentlicht in:Analytical biochemistry 1987-11, Vol.166 (2), p.267-275
Hauptverfasser: Johansson, G., Öfverstedt, L.-G., Hjertén, S.
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container_title Analytical biochemistry
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creator Johansson, G.
Öfverstedt, L.-G.
Hjertén, S.
description This paper describes the separation of proteins by displacement electrophoresis on columns packed with cellulose powder as a stabilizing medium. Cellulose has virtually no molecularsieving 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.
doi_str_mv 10.1016/0003-2697(87)90574-4
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subjects Analytical biochemistry: general aspects, technics, instrumentation
Analytical, structural and metabolic biochemistry
Biological and medical sciences
Blood Protein Electrophoresis
Cellulose
cellulose powder column
continuous elution
counter flow
displacement electrophoresis
Electrophoresis - instrumentation
Electrophoresis, Polyacrylamide Gel
Erythrocytes - analysis
Fundamental and applied biological sciences. Psychology
gel electrophoresis
isotachophoresis
protein aggregation
proteins
Proteins - analysis
spacer techniques
title Preparative displacement electrophoresis (isotachophoresis) of proteins on cellulose columns
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