Capillary zone electrophoretic resolution of recombinant human bone morphogenetic protein 2 glycoforms an investigation into the separation mechanisms for an exquisite separation

Yim, Kalvin; Abrams, Joan; Hsu, Amy

doi:10.1016/0021-9673(95)00742-6

Cited by 24 publications

(12 citation statements)

References 20 publications

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“…CZE separation was performed according to their number of glycosyl moieties linked to the polypeptide chain of b-conglycinin. CZE has been previously employed as a successful technique for studying intact glycoproteins and glycopeptides (Balaguer et al, 2006;Berkowitz et al, 2005;Weber, Kornfelt, Klausen, & Lunte, 1995;Yim, Abrams, & Hsu, 1995).…”

Section: Effective Deglycosylation Of B-conglycinin Soy Allergenmentioning

confidence: 99%

Carbohydrate moieties on the in vitro immunoreactivity of soy β-conglycinin

Amigo‐Benavent

Athanasopoulos

Ferranti

et al. 2009

Food Research International

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Section: Effective Deglycosylation Of B-conglycinin Soy Allergenmentioning

confidence: 99%

Carbohydrate moieties on the in vitro immunoreactivity of soy β-conglycinin

Amigo‐Benavent

Athanasopoulos

Ferranti

et al. 2009

Food Research International

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“…Sialic acid would render the glycoform less positively charged, and then it would migrate earlier in this cationic-coated capillary. However, CZE separations of glycoforms that differ only in the number of mannose residues and thus in size have been shown for other glycoprotein [54]. About 22% of recombinant human VEGF 165 corresponds to the glycosidic moiety [15]; thus the larger size of the glycoforms compared with the non-glycosylated forms would also contribute to their lower effective mobility opposite to the EOF and thus to a shorter t m for the glycosylated than for the non-glycosylated forms.…”

Section: Assignment Accuracy Of Peaks Of Vegfmentioning

confidence: 95%

Development of CE methods to analyze potential components of the angiogenic glycoprotein vascular endothelial growth factor 165

et al. 2009

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The vascular endothelial growth factor 165 (VEGF165) is the predominant form of the complex VEGF family. This glycoprotein has, among others, an angiogenic effect in many physiological and pathological events. For this reason, its roles as a biomarker and as a therapeutic drug have been considered. However, very little is known about the existence of different forms of VEGF165 arising from glycosylation and other potential PTMs. This aspect is crucial because it is known that for other glycoproteins the ratio between these isoforms actually acts as a biomarker for certain diseases and other physiological states. In addition, for therapeutic use of glycoproteins it is known that the biological activity may differ for the various isoforms. In this work CE methods to separate up to seven peaks without baseline resolution containing various forms of VEGF165 are developed. Using a computer program previously developed in‐house peak assignment could be performed with accuracy close to 100%. In this way, comparison between recombinant human VEGF165 expressed in insect cells, which is a glycosylating system, and in Escherichia coli cells, which are unable of performing glycosylation of proteins, has been possible. The methods developed, besides providing information about the existence of several forms of VEGF165, mean a starting point that permits the study of the role of VEGF165 as a potential biomarker of different diseases and physiological processes and to perform quality control of the recombinant drug during manufacturing. To the best of our knowledge this is the first time that CE methods for VEGF165 have been developed.

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“…Before 2000 there were only two papers published reporting on separation of the glycoforms of rhBMP-2 by capillary electrophoresis coupled to MS. (Yeung et al, 1997;Yim et al, 1995) From 2000 onward, there is a continuum of scientific research on the BMPs using a mass spectrometry. There are two main approaches to the research in the field: (a) isolation and identification of a BMP or related molecule from different biological sources; and (b) proteomic study of a system perturbed using a BMP or related molecule.…”

Section: Bone Morphogenetic Proteins By Mass Spectrometrymentioning

confidence: 99%