Optimization of a nano‐enzymatic reactor for on‐line tryptic digestion of polypeptide conjugates by capillary electrophoresis

摘要: This work aims at studying the optimization of an on-line capillary electrophoresis (CE)-based tryptic digestion methodology for analysis therapeutic polypeptides (PP). With this methodology, a mixture surrogate peptide fragments and amino acid were produced by trypsin cleavage (enzymatic digestion) subsequently analyzed using same capillary. The resulting automation all steps such as injection, mixing, incubation, separation detection minimizes possible errors saves experimental time. In paper, we first study differents parameters influencing PP inside (plug length, reactant concentration, incubation time, diffusion electrophoretic plugs mixing). second part, conditions generated hydrolysis products (nature, pH ionic strength (I) background electrolyte (BGE)) is described. Using optimized conditions, excellent repeatability was obtained in terms (migration times) proteolysis (number from enzymatic corresponding amounts) demonstrating robustness proposed methodology.