Direct quality control of glycoengineered erythropoietin variants
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Direct quality control of glycoengineered erythropoietin variants. / Čaval, Tomislav; Tian, Weihua; Yang, Zhang; Clausen, Henrik; Heck, Albert J.R.
I: Nature Communications, Bind 9, 3342, 2018.Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › fagfællebedømt
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T1 - Direct quality control of glycoengineered erythropoietin variants
AU - Čaval, Tomislav
AU - Tian, Weihua
AU - Yang, Zhang
AU - Clausen, Henrik
AU - Heck, Albert J.R.
PY - 2018
Y1 - 2018
N2 - Recombinant production of glycoprotein therapeutics like erythropoietin (EPO) in mammalian CHO cells rely on the heterogeneous N-glycosylation capacity of the cell. Recently, approaches for engineering the glycosylation capacities of mammalian cells for custom designed glycoforms have been developed. With these opportunities there is an increasing need for fast, sensitive, and global analysis of the glycoproteoform landscape produced to evaluate homogeneity and consistency. Here we use high-resolution native mass spectrometry to measure the glycoproteoform profile of 24 glycoengineered variants of EPO. Based on the unique mass and intensity profiles of each variant, we classify them according to similarities in glycosylation profiles. The classification distinguishes EPO variants with varying levels of glycan branchingand sialylation, which are crucial parameters in biotherapeutic efficacy. We propose that our methods could be of great benefit in the characterization of other glycosylated biopharmaceuticals, ranging from the initial clonal selection to batch-to-batch controls, and the assessment of similarity between biosimilar/biobetter products.
AB - Recombinant production of glycoprotein therapeutics like erythropoietin (EPO) in mammalian CHO cells rely on the heterogeneous N-glycosylation capacity of the cell. Recently, approaches for engineering the glycosylation capacities of mammalian cells for custom designed glycoforms have been developed. With these opportunities there is an increasing need for fast, sensitive, and global analysis of the glycoproteoform landscape produced to evaluate homogeneity and consistency. Here we use high-resolution native mass spectrometry to measure the glycoproteoform profile of 24 glycoengineered variants of EPO. Based on the unique mass and intensity profiles of each variant, we classify them according to similarities in glycosylation profiles. The classification distinguishes EPO variants with varying levels of glycan branchingand sialylation, which are crucial parameters in biotherapeutic efficacy. We propose that our methods could be of great benefit in the characterization of other glycosylated biopharmaceuticals, ranging from the initial clonal selection to batch-to-batch controls, and the assessment of similarity between biosimilar/biobetter products.
U2 - 10.1038/s41467-018-05536-3
DO - 10.1038/s41467-018-05536-3
M3 - Journal article
C2 - 30131559
AN - SCOPUS:85051840279
VL - 9
JO - Nature Communications
JF - Nature Communications
SN - 2041-1723
M1 - 3342
ER -
ID: 209061727