Evaluation of the electroosmotic medium pump system for preparative disk gel electrophoresis

This paper describes an improved electroosmotic elution system for preparat ive sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) ba sed on the epochal idea of H. V. Tan ct al. (Nucleic Acids Res. 1988, 16, 1 921-1930). In this elution system, a semipermeable membrane, mounted under the gel terminal end, works as the elution pump as web as the partition of the elution chamber. We refer to this system as the "electroosmotic medium pump system." Operation of the constructed apparatus (3.6 cm i.d. disk gel column) and resolution of the protein bands were examined by separation of the model protein mixture (bovine serum albumin (ESA), ovalbumin, bovine ca rbonic anhydrase, soybean trypsin inhibitor) and purification of the membra ne protein, dipeptidyl peptidase IV (DPP TV). The Spectra/Por 7 dialysis me mbrane provided a better how profile for the elution buffer. The four model proteins of the protein mixture mere able to be completely separated from each other and recovered without dilution. The maximum protein concentratio n of eluate achieved was 93 mg/ml, when applying a single component, BSA fr action V, as a sample. Furthermore, the multifunctional ectoenzyme, DPP IV, was purified in a single step. (C) 2001 Academic Press.