TY - JOUR

T1 - The GalNAc-type O-glycoproteome of CHO cells characterized by the SimpleCell strategy

AU - Yang, Zhang

AU - Halim, Adnan

AU - Narimatsu, Yoshiki

AU - Joshi, Hiren Jitendra

AU - Steentoft, Catharina

AU - Schjoldager, Katrine Ter-Borch Gram

AU - Schulz, Morten Alder

AU - Sealover, Natalie R

AU - Kayser, Kevin J

AU - Bennett, Eric Paul

AU - Levery, Steven B

AU - Vakhrushev, Sergey Y

AU - Clausen, Henrik

N1 - Copyright © 2014, The American Society for Biochemistry and Molecular Biology.

PY - 2014/12

Y1 - 2014/12

N2 - The Chinese hamster ovary cell (CHO) is the major host cell factory for recombinant production of biological therapeutics primarily due to 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. While 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 (1,548 O-glycosites) in cell lysates and secretomes providing the first comprehensive insight into the O-glycosylation capacity of CHO (https://glycomics.ku.dk/o-glycoproteome_db/).

AB - The Chinese hamster ovary cell (CHO) is the major host cell factory for recombinant production of biological therapeutics primarily due to 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. While 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 (1,548 O-glycosites) in cell lysates and secretomes providing the first comprehensive insight into the O-glycosylation capacity of CHO (https://glycomics.ku.dk/o-glycoproteome_db/).

U2 - 10.1074/mcp.M114.041541

DO - 10.1074/mcp.M114.041541

M3 - Journal article

C2 - 25092905

VL - 13

SP - 3224

EP - 3235

JO - Molecular and Cellular Proteomics

JF - Molecular and Cellular Proteomics

SN - 1535-9476

IS - 12

ER -