The GalNAc-type O-Glycoproteome of CHO Cells Characterized by the SimpleCell Strategy

Overview

Journal Mol Cell Proteomics

Specialties Biochemistry
Cell Biology
Molecular Biology

Date 2014 Aug 6

PMID 25092905

Citations 42

Authors

Zhang Yang

Adnan Halim

Yoshiki Narimatsu

Hiren Jitendra Joshi

Catharina Steentoft

Katrine Ter-Borch Gram Schjoldager

Morten Alder Schulz

Natalie R Sealover

Kevin J Kayser

Eric Paul Bennett

Steven B Levery

Sergey Y Vakhrushev

Henrik Clausen

Affiliations

Soon will be listed here.

Abstract

The Chinese hamster ovary cell (CHO) is the major host cell factory for recombinant production of biological therapeutics primarily because of its "human-like" glycosylation features. CHO is used for production of several O-glycoprotein therapeutics including erythropoietin, coagulation factors, and chimeric receptor IgG1-Fc-fusion proteins, however, some O-glycoproteins are not produced efficiently in CHO. We have previously shown that the capacity for O-glycosylation of proteins can be one limiting parameter for production of active proteins in CHO. Although the capacity of CHO for biosynthesis of glycan structures (glycostructures) on glycoproteins are well established, our knowledge of the capacity of CHO cells for attaching GalNAc-type O-glycans to proteins (glycosites) is minimal. This type of O-glycosylation is one of the most abundant forms of glycosylation, and it is differentially regulated in cells by expression of a subset of homologous polypeptide GalNAc-transferases. Here, we have genetically engineered CHO cells to produce homogeneous truncated O-glycans, so-called SimpleCells, which enabled lectin enrichment of O-glycoproteins and characterization of the O-glycoproteome. We identified 738 O-glycoproteins (1548 O-glycosites) in cell lysates and secretomes providing the first comprehensive insight into the O-glycosylation capacity of CHO (http://glycomics.ku.dk/o-glycoproteome_db/).

Citing Articles

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