Isotachophoretic analysis of peptides. Selection of electrolyte systems and determination of purity.
Capillary isotachophoresis (ITP) was applied to the qualitative and quantitative analysis of both natural and synthetic oligo- and polypeptides. Based on the mathematical model of acid-base equilibria for a general ampholyte, a procedure and a computer program for the calculation of the pH dependence of the effective and specific charge and effective mobility of peptides with known amino acid sequence were developed which allow the selection of electrolyte systems for peptide isotachophoretic analysis to be rationalized. Basic peptides (bovine pancreatic trypsin inhibitor, bull seminal isoinhibitors of trypsin, arginine vasopressin and adamantylamide-alanylisoglutamine) were analysed with a cationic ITP system at acidic pH. Neutral and acidic peptides (insulin, proinsulin, bull seminal isoinhibitors of trypsin, cow colostrum isoinhibitors of trypsin) were analysed with an anionic ITP system, mostly at alkaline pH. Peptide purity (electrophoretic homogeneity) was determined from the ITP degree of purity defined by a peptide itself and the zone length ratio of its admixtures. Enrichment of peptide in the sample during the purification procedure was measured by its zone length relative to unit mass of the amount of sample analysed.
Kasicka
V
V
Institute of Organic Chemistry and Biochemistry, Czechoslovak Academy of Sciences, Prague.
Prusík
Z
Z
D000665:Ampholyte Mixtures; D000818:Animals; D002417:Cattle; D004586:Electrophoresis / Q000379:methods*; D006466:Hemopexin / Q000032:analysis; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007328:Insulin / Q000032:analysis; D010455:Peptides / Q000032:analysis*; D013056:Spectrophotometry, Ultraviolet; D013552:Swine; D014361:Trypsin Inhibitors / Q000032:analysis
D000665:Ampholyte Mixtures; D007328:Insulin; D010455:Peptides; D014361:Trypsin Inhibitors; D006466:Hemopexin