Immunochromatographic removal of albumin in erythropoietin biopharmaceutical formulations for its analysis by capillary electrophoresis

Lara-Quintanar, Pilar; Lacunza, Izaskun; Sanz, J.; Diez-Masa, Jose Carlos; Frutos, Mercedes de

doi:10.1016/j.chroma.2006.07.079

Cited by 25 publications

(8 citation statements)

References 25 publications

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“…The detailed description of the computer program and the algorithm for peak assignment has been previously published [27]. The applicability of this program for other glycoproteins has been demonstrated in previous works [28][29][30]. Further details about performing assignment are available in the Supporting Information.…”

Section: Assignment Of Vegf 165 Peaks: Computer Program and Migrationmentioning

confidence: 97%

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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Section: Assignment Of Vegf 165 Peaks: Computer Program and Migrationmentioning

confidence: 97%

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

et al. 2009

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“…Albumin is used as rhEPO stabilizer and both were good separated by CE however without good sensitivity [37]. A trial to increase sensitivity was done by immunochromatographic removal of albumin in erythropoietin biopharmaceutical formulations for its analysis by CE [38]. However, this method was complex, expensive and time consuming.…”

Section: Introductionmentioning

confidence: 99%

The use of laser-induced fluorescence or ultraviolet detectors for sensitive and selective analysis of tobramycin or erythropoietin in complex samples

Ahmed

Ebeid²

2015

Spectrochimica Acta Part A: Molecular and Biomolecular Spectros

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“…Reversed-phase liquid chromatography (RP-LC) exploits the hydrophobic properties of the biomolecules in the separation process and offers a high level of accuracy and sensitivity for the analysis of closely related protein variants which may have reduced activity and altered immunogenicity. 21,22 A linear gradient RP-LC method using a C 4 column and UV detection at 280 nm was validated for the analysis of rhEPO in biopharmaceutical formulations and the results compared to the bioassay show differences that are 11.2% higher for the LC method. 23 A size-exclusion (SE-LC) method was validated for the determination of rhEPO in formulations without serum albumin, using a BioSep-SEC-S 2000 column and detection at 214 nm.…”

Section: Introductionmentioning

confidence: 99%

Assessment of recombinant human erythropoietin by alternative bioassay and its correlation with liquid chromatography methods

et al. 2013

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Biotechnology-derived erythropoietin is a sialoglycoprotein, which stimulates erythropoiesis, and it is clinically used for the treatment of anaemia related to chronic renal failure. N-Acetylneuraminic acid content was quantified by a reversed-phase liquid chromatography method with fluorescence detection giving values higher than 108.74 ng mg À1 . An alternative in vitro TF-1 cell proliferation bioassay was studied showing a lower mean difference of the estimated potency of 2.67%, compared to the normocythaemic mice bioassay, with non-significant differences (p > 0.05). Biopharmaceutical products were also analyzed by validated reversed-phase and size-exclusion liquid chromatography methods and compared to the in vivo bioassay, showing lower mean differences of the content/potencies of 2.11 and 1.21%, respectively. Higher molecular mass forms and deamidated/sulphoxide forms showed mean bioactivities reduced to about 10%, for both. The TF-1 cell culture assay in conjunction with the determination of sialic acids represents an advance that can be correlated with the normocythaemic mice bioassay and the physicochemical methods, allowing for the establishment of alternative methods, which can be applied to monitor the stability, quality control, and thereby ensuring the therapeutic efficacy of the biological medicine.

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Immunochromatographic removal of albumin in erythropoietin biopharmaceutical formulations for its analysis by capillary electrophoresis

Cited by 25 publications

References 25 publications

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

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

The use of laser-induced fluorescence or ultraviolet detectors for sensitive and selective analysis of tobramycin or erythropoietin in complex samples

Assessment of recombinant human erythropoietin by alternative bioassay and its correlation with liquid chromatography methods

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