title: Separation of peptides dissolved in a sodium dodecyl sulfate solution by reversed-phase liquid chromatography: removal of sodium dodecyl sulfate from peptides using an ion-exchange precolumn.
abstract: Separation of peptides by reversed-phase liquid chromatography is significantly affected by sodium dodecyl sulfate (SDS) in the sample solution. The strongly acidic group of SDS binds to the reversed-phase column where it serves as an ion exchanger and retards the elution of peptides. By using a DEAE precolumn connected in series to a reversed-phase column, the interference of SDS in the separation of peptides by reversed-phase chromatography can be significantly diminished. This simple method is applicable to the separation of peptide mixtures obtained by digestion of proteins extracted from SDS-polyacrylamide gels. Peptide production with some proteases in the presence of SDS was examined using the present method. Lysylendopeptidase was suitable for digestion in the presence of SDS, but V8 protease was not.
authors.affiliation: Department of Molecular Biology, Tokyo Metropolitan Institute of Medical Science, Japan.
authors.forename: H
authors.initials: H
authors.lastname: Kawasaki
authors.forename: K
authors.initials: K
authors.lastname: Suzuki
mesh_terms: D000595:Amino Acid Sequence; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D004983:Ethanolamines; D008969:Molecular Sequence Data; D010455:Peptides / Q000302:isolation & purification* / Q000378:metabolism; D011108:Polymers; D012697:Serine Endopeptidases / Q000378:metabolism; D012967:Sodium Dodecyl Sulfate
chemical_list: C064803:DEAE-Toyopearl 650S; D004983:Ethanolamines; D010455:Peptides; D011108:Polymers; D012967:Sodium Dodecyl Sulfate; D012697:Serine Endopeptidases; C028344:glutamyl endopeptidase; C036813:lysyl endopeptidase