Performance evaluation of capillary surface treatments for peptide mapping by capillary zone electrophoresis
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The performance of different capillary coatings and their ability to reduce peptide/wall interactions in capillary zone electrophoresis (CZE) was evaluated through the separations of standard peptides with PM ranging from 720 to 3500 and pl ranging from 3.0 to 11.5. The study was performed in acidic buffer with untreated fused silica capillary, poly(vinylalcohol) (PVA) modified capillary (generated in our laboratory by immobilization with thermal treatment of a PVA solution) and polybrene coated capillary. PVA and polybrene modified capillaries were shown to produce higher reproducibilities for migration times and superior efficiencies in comparison to the uncoated capillary. Peak efficiencies of acidic peptides were 300000 to 650000 (N/m) with polybrene capillary coating while PVA coating produced high efficiencies (N/m>330000) for the most basic peptides. An efficient peptide mapping of the β-lactoglobulin was obtained in less than 20 min using the polybrene coating. Attempts to separate the peptide digest from a deglycosylated form of the rgp 160, a very complex and heterogeneous HIV envelope glycoprotein, were not fully satisfactory but the results demonstrate the potential of capillary surface treatment in producing peptide maps.
Key WordsCapillary zone electrophoresis Capillary coating Peptide mapping HIV envelope glycoprotein
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