Capillary electrophoretic separation of recombinant granulocyte-colony-stimulating factor glycoforms

Watson, Eric; Yao, Fei-Long

doi:10.1016/0021-9673(93)80485-q

Cited by 28 publications

(5 citation statements)

References 10 publications

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“…Partial or a nearly complete resolution of glycoforms through HPCE has now been demonstrated with a variety of natural glycoproteins: ribonuclease B, − bovine fetuin, ovalbumin, ,− α 1 -acid glycoprotein, ,, horseradish peroxidase, , pepsin, human chorionic gonadotrophin, and hirudin . In addition, several recombinant glycoproteins of medical importance, including the recombinant human blood coagulation factor VII, soluble T4 receptor protein, human bone morphogenic protein-2, human granulocyte-macrophage colony stimulating factor, and both natural and recombinant interleukin-2, were also analyzed by CE. While some of these biomolecules were used as “model glycoproteins” to demonstrate the technique's effectiveness and establish the critical separation parameters, it is apparently feasible to study unknown glycoproteins so long as their complexity is not extraordinarily high.…”

Section: Glycoform Separations By Capillary Electrophoresismentioning

confidence: 99%

Structural Investigations of Glycoconjugates at High Sensitivity

2002

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Section: Glycoform Separations By Capillary Electrophoresismentioning

confidence: 99%

Structural Investigations of Glycoconjugates at High Sensitivity

2002

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“…A set of different microscale techniques of CE exists for analyzing recombinant glycoproteins. CZE is the general method in this field for analyzing recombinant protein variants, such as human EPO [139,140], human chorionic gonadotropin [141], human tissue plasminogen activator [142], human granulocyte-macorphage colony stimulating factor [143], and human blood coagulation factor VII [144]. All of these examples were performed in uncoated silicafused capillaries.…”

Section: Capillary Electrophoresismentioning

confidence: 99%

Chromatographic and electrophoretic characterization of protein variants

Ahrer

Jungbauer

2006

Journal of Chromatography B

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“…Watson and Yao were able to separate rhuEPO into at least six glycoforms using an uncoated fused-silica capillary with a tricine buffer at pH 6.2, containing 1,4-diaminobutane (DAB) to reduce EOF, and 7 M urea for resolution and peak shape [43]. DAB was also used as a buffer additive to resolve recombinant granulocyte colony stimulating factor (rGCSF, [44]) and factor VIla glycoforms [45] and was therefore useful for resolving sialic acid-containing variants. Yim et al [46] used a phosphate buffer system at pH 2.5, with no additives, to Original analyze nine glycoforms of recombinant human bone morphogenic protein 2 (rhBMP2).…”

Section: Reviews Of Ce Used For the Analysis Of Recombinant Proteinsmentioning

confidence: 99%

Capillary Electrophoresis in the Development of Recombinant Protein Biopharmaceuticals

Chen¹,

Canova-Davis²

2001

CE in Biotechnology: Practical Applications for Protein and Peptide Analyses

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The understanding of the physicochemical properties of proteins in solution is required to understand the separations taking place in the capillary. For example, during isoelectric focusing, it is important to ensure the lack of protein precipitation during fOCUSing and mobilization otherwise artifacts occur and the true resolution of charge variants is not achieved. Similarly, buffer conditions, including additives, composition of buffer, and pH can influence the resolution of product variants. With diligent efforts one can achieve desired separations. As knowledge of buffer conditions and use of these techniques increase, the methods will be improved.MECC can be useful for the characterization of recombinant proteins and monoclonal antibodies. However, the relatively long analyte migration times may be a deterrent for routine use in quality control situations. Pharmaceutical companies will employ new techniques which provide advantages over current technology. CE has advantages in certain applications. Applications which use slab gel techniques, IEF and SDS-PAGE can be replaced by the CE methods for formulation development, characterization of charge isoforms, and for purity determinations. CZE and clEF can be used for rapid identity tests which do not require peptide mapping. CZE has a tremendous advantage in analyzing basic proteins, and CZE and CE-SDS can be used to quantify product in the presence of interfering excipients. It is the responsibility of the analytical chemist to determine applications which are appropriate for CEo It is apparent that these are currently being defined for the biotechnology industry, and the usage of CE should continue to increase.

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Capillary electrophoretic separation of recombinant granulocyte-colony-stimulating factor glycoforms

Cited by 28 publications

References 10 publications

Structural Investigations of Glycoconjugates at High Sensitivity

Structural Investigations of Glycoconjugates at High Sensitivity

Chromatographic and electrophoretic characterization of protein variants

Capillary Electrophoresis in the Development of Recombinant Protein Biopharmaceuticals

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