CHARACTERIZATION OF GLYCOPEPTIDES FROM RECOMBINANT COAGULATION-FACTORVIIA BY HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY AND CAPILLARY ZONE ELECTROPHORESIS USING ULTRAVIOLET AND PULSED ELECTROCHEMICAL DETECTION

Four glycopeptide (GP) fractions containing glycosylated Asn 322 were isolated from a tryptic digest of recombinant coagulation factor VII b y reversed-phase-HPLC (RP-HPLC). Characterization of the GPs by enzyma tic desialylation and RP-HPLC as well as by enzymatic deglycosylation, RP-HPLC, and high-pH anion-exchange chromatography indicated that the four GPs consisted of the same decapeptide but with O, 1, 2, or 4 res idues of sialic acid. In comparison to HPLC, capillary zone electropho resis (CZE) using uv and pulsed electrochemical detection (FED) afford ed improved separation of GPs from each other and from contaminants. C ZE-uv and CZE-PED of the desialylated GPs and deglycosylated GrPs corr oborated the results obtained with the chromatographic methods. (C) 19 95 Academic Press, Inc.