A sialylation-sensitive cell-based in vitro bioassay for erythropoietin; incorporation of the galactose-binding Erythrina crista-galli lectin

Liefooghe, Emily C.; Tiplady, Richard; Gerson, Peter; Lloyd, Pauline; Heath, Alan; Bristow, Adrian

doi:10.1016/j.biologicals.2005.05.001

Cited by 10 publications

(6 citation statements)

References 18 publications

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“…Particularly, native MS is extremely powerful at quantifying the distinct PTM patterns of glycoproteins with heavy sialylation. These monosaccharides are substantially important for the biological activities and the in vivo circulatory half-life of many biopharmaceutical products, such as rhEPO4445, yet are very challenging for quantification because they introduce ionization bias to the sialylated glycopeptide and quite fragile during sample preparation.…”

Section: Discussionmentioning

confidence: 99%

Hybrid mass spectrometry approaches in glycoprotein analysis and their usage in scoring biosimilarity

et al. 2016

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Many biopharmaceutical products exhibit extensive structural micro-heterogeneity due to an array of co-occurring post-translational modifications. These modifications often effect the functionality of the product and therefore need to be characterized in detail. Here, we present an integrative approach, combining two advanced mass spectrometry-based methods, high-resolution native mass spectrometry and middle-down proteomics, to analyse this micro-heterogeneity. Taking human erythropoietin and the human plasma properdin as model systems, we demonstrate that this strategy bridges the gap between peptide- and protein-based mass spectrometry platforms, providing the most complete profiling of glycoproteins. Integration of the two methods enabled the discovery of three undescribed C-glycosylation sites on properdin, and revealed in addition unexpected heterogeneity in occupancies of C-mannosylation. Furthermore, using various sources of erythropoietin we define and demonstrate the usage of a biosimilarity score to quantitatively assess structural similarity, which would also be beneficial for profiling other therapeutic proteins and even plasma protein biomarkers.

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Section: Discussionmentioning

confidence: 99%

Hybrid mass spectrometry approaches in glycoprotein analysis and their usage in scoring biosimilarity

et al. 2016

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“…This impacted the rules of the innovation process by introducing a new barrier to the use of animal-free methods. (Egrie et al, 1987) + − − Indirect Cultured cells (Krystal et al, 1981) + + − Indirect Western method (Hammerling et al, 1996) + − − Indirect Sialylation-sensitive cell method (Liefooghe et al, 2005) + + ± Indirect RP -LC method (Barth et al, 2007) − − + Indirect MALDI TO F MS method (Llop et al, 2008) − − + Indirect CZE method (Zhang et al, 2009) − − + Indirect cIE F method (Cifuentes et al, 1999) − − + Indirect ⁎ This list of criteria is not exhaustive and not prioritized.…”

Section: Deadlocked Innovation Due To Collective Inactionmentioning

confidence: 99%

How institutional logics hamper innovation: The case of animal testing

Kooijman

Hekkert

Meer

et al. 2017

Technological Forecasting and Social Change

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For radical innovation to become successful the substitution of established practices are essential. Nevertheless, in the innovation literature novelty is often at the center and only little attention is paid to the influence of established technologies and underlying routines. This paper aims to contribute to this gap by increasing the understanding about the effect of persistence of established practices on the innovation process. We do this by using a framework that combines the Technological Innovation System approach with an analysis of the institutional logics reinforcing the established practice. The studied case concerns the innovation process to animal-free medicine development. Despite the fact that the substitution of animal tests is called for since the 1980s and animalfree methods are available, animal tests are still being used in medicine development. This study shows that adding institutional logics to the innovation systems analysis creates a much better understanding of the speed and direction of radical innovation.

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“…The normocythaemic mouse bioassay has been performed with either single or multiple daily injection protocols on normal ani-mals, with blood collection on the 5th day and reticulocyte counting by flow cytometry (6). An in vitro sialylation-sensitive bioassay that makes use of the human AS-E2 EPO-dependent cell line has been shown to give results which correlate with the in vivo polycythaemic mouse bioassay (7). In another in vitro cell culture assay, which employs the well-characterised factor-dependent human erythroleukaemia cell line, TF-1 (ATCC CRL-2003), it was found that cell proliferation was dependent on EPO (8).…”

Section: Introductionmentioning

confidence: 99%

Evaluation of an In Vitro Cell Culture Assay for the Potency Assessment of Recombinant Human Erythropoietin

et al. 2016

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Recombinant human erythropoietin is a sialoglycoprotein that stimulates erythropoiesis. To assess potency of human erythropoietin produced by recombinant technology, we investigated an in vitro TF-1 cell proliferation assay, which was applied in conjunction with a reversed-phase liquid chromatography method for the determination of the content of sialic acids. The results obtained, which were higher than 126.8ng/μg, were compared with those obtained with the in vivo normocythaemic mouse bioassay. The in vitro assay resulted in a non-significant lower mean difference of the estimated potencies (0.61% ± 0.026, p > 0.05). The use of this combination of methods represents an advance toward the establishment of alternative in vitro approaches, in the context of the Three Rs, for the potency assessment of biotechnology-derived medicines.

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A sialylation-sensitive cell-based in vitro bioassay for erythropoietin; incorporation of the galactose-binding Erythrina crista-galli lectin

Cited by 10 publications

References 18 publications

Hybrid mass spectrometry approaches in glycoprotein analysis and their usage in scoring biosimilarity

Hybrid mass spectrometry approaches in glycoprotein analysis and their usage in scoring biosimilarity

How institutional logics hamper innovation: The case of animal testing

Evaluation of an In Vitro Cell Culture Assay for the Potency Assessment of Recombinant Human Erythropoietin

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