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Application of capillary electrophoresis, liquid chromatography, electrospray-mass spectrometry and matrix-assisted laser desorption/ionization - time of flight - mass spectrometry to the characterization of recombinant human erythropoietin.
Electrophoresis. 1998 Oct; 19(13):2348-55.E

Abstract

High performance capillary electrophoresis (HPCE), high performance liquid chromatography (HPLC), matrix-assisted laser desorption/ionization - time of flight - mass spectrometry (MALDI-TOF-MS), on-line CE-electrospray ionization-mass spectrometry (CE-ESI-MS) and on-line LC-ESI-MS have been employed to characterize a heterogeneous glycoprotein, recombinant human erythropoietin (rHuEPO) expressed from Chinese hamster ovary (CHO) cells. The analysis was demonstrated through two specific levels of detail: the intact protein and tryptic digests of the protein. Six glycoforms of rHuEPO were separated by HPCE; seventeen tryptic fragments in a total of 21 nonglycosylated and glycosylated peptides were characterized; the O-linked glycopeptides were analyzed directly by CE-ESI-MS and LC-ESI-MS. In particular, four glycans of O-acetylation of sialic acid were identified in the O-linked glycosylated fragments. The molecular weight of rHuEPO was accurately determined by MALDI-TOF-MS.

Authors+Show Affiliations

Department of Chemistry, Tsinghua University, Beijing, PRC.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

9788319

Citation

Zhou, G H., et al. "Application of Capillary Electrophoresis, Liquid Chromatography, Electrospray-mass Spectrometry and Matrix-assisted Laser Desorption/ionization - Time of Flight - Mass Spectrometry to the Characterization of Recombinant Human Erythropoietin." Electrophoresis, vol. 19, no. 13, 1998, pp. 2348-55.
Zhou GH, Luo GA, Zhou Y, et al. Application of capillary electrophoresis, liquid chromatography, electrospray-mass spectrometry and matrix-assisted laser desorption/ionization - time of flight - mass spectrometry to the characterization of recombinant human erythropoietin. Electrophoresis. 1998;19(13):2348-55.
Zhou, G. H., Luo, G. A., Zhou, Y., Zhou, K. Y., Zhang, X. D., & Huang, L. Q. (1998). Application of capillary electrophoresis, liquid chromatography, electrospray-mass spectrometry and matrix-assisted laser desorption/ionization - time of flight - mass spectrometry to the characterization of recombinant human erythropoietin. Electrophoresis, 19(13), 2348-55.
Zhou GH, et al. Application of Capillary Electrophoresis, Liquid Chromatography, Electrospray-mass Spectrometry and Matrix-assisted Laser Desorption/ionization - Time of Flight - Mass Spectrometry to the Characterization of Recombinant Human Erythropoietin. Electrophoresis. 1998;19(13):2348-55. PubMed PMID: 9788319.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Application of capillary electrophoresis, liquid chromatography, electrospray-mass spectrometry and matrix-assisted laser desorption/ionization - time of flight - mass spectrometry to the characterization of recombinant human erythropoietin. AU - Zhou,G H, AU - Luo,G A, AU - Zhou,Y, AU - Zhou,K Y, AU - Zhang,X D, AU - Huang,L Q, PY - 1998/10/27/pubmed PY - 1998/10/27/medline PY - 1998/10/27/entrez SP - 2348 EP - 55 JF - Electrophoresis JO - Electrophoresis VL - 19 IS - 13 N2 - High performance capillary electrophoresis (HPCE), high performance liquid chromatography (HPLC), matrix-assisted laser desorption/ionization - time of flight - mass spectrometry (MALDI-TOF-MS), on-line CE-electrospray ionization-mass spectrometry (CE-ESI-MS) and on-line LC-ESI-MS have been employed to characterize a heterogeneous glycoprotein, recombinant human erythropoietin (rHuEPO) expressed from Chinese hamster ovary (CHO) cells. The analysis was demonstrated through two specific levels of detail: the intact protein and tryptic digests of the protein. Six glycoforms of rHuEPO were separated by HPCE; seventeen tryptic fragments in a total of 21 nonglycosylated and glycosylated peptides were characterized; the O-linked glycopeptides were analyzed directly by CE-ESI-MS and LC-ESI-MS. In particular, four glycans of O-acetylation of sialic acid were identified in the O-linked glycosylated fragments. The molecular weight of rHuEPO was accurately determined by MALDI-TOF-MS. SN - 0173-0835 UR - https://www.unboundmedicine.com/medline/citation/9788319/Application_of_capillary_electrophoresis_liquid_chromatography_electrospray_mass_spectrometry_and_matrix_assisted_laser_desorption/ionization___time_of_flight___mass_spectrometry_to_the_characterization_of_recombinant_human_erythropoietin_ L2 - https://doi.org/10.1002/elps.1150191315 DB - PRIME DP - Unbound Medicine ER -