O-glycans and O-glycosylation sites of recombinant human GM-CSF derived from suspension-cultured rice cells, and their structural role

Kim, Jihye; Park, Heajin; Park, Byung Tae; Hwang, Hye Seong; Kim, Jae Il; Kim, Dae Kyong; Kim, Ha Hyung

doi:10.1016/j.bbrc.2016.09.057

Cited by 10 publications

(5 citation statements)

References 35 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“… 26 , 27 , 28 As many therapeutic proteins, such as fusion proteins etanercept, abatacept, atacicept, hormone erythropoietin (EPO), recombinant coagulation factor VIII, granulocyte macrophage colony stimulating factor, and others are both O-glycosylated and N-glycosylated. 19 , 29 , 30 , 31 , 32 , 33 , 34 The role of N-glycosylation in quality and safety of those protein drugs are well known. However, the effects of O-glycosylation on critical quality attributes of these therapeutic proteins are currently largely uninvestigated.…”

Section: Discussionmentioning

confidence: 99%

An etanercept O-glycovariant with enhanced potency

Biel

Faison

Matthews

et al. 2022

Molecular Therapy - Methods & Clinical Development

View full text Add to dashboard Cite

Section: Discussionmentioning

confidence: 99%

An etanercept O-glycovariant with enhanced potency

Biel

Faison

Matthews

et al. 2022

Molecular Therapy - Methods & Clinical Development

View full text Add to dashboard Cite

“…As a consequence, plant-specific O-glycosylation of Ser, as well as hydroxylation and O-glycosylation of Pro residues, have been reported on biopharmaceuticals produced in plants because peptide motifs within these recombinant protein sequence have been recognized by the plant Golgi glycosyltransferases involved in HRGP biosynthesis. This gives rise to unwanted modifications that could potentially impair the protein functionality or induce immune responses [23,29,30]. Unexpected arabinose residues have also been reported on N-glycans of biopharmaceuticals expressed in glyco-humanized plants.…”

Section: Discussionmentioning

confidence: 99%

Investigation of the N-Glycosylation of the SARS-CoV-2 S Protein Contained in VLPs Produced in Nicotiana benthamiana

et al. 2022

View full text Add to dashboard Cite

The emergence of the SARS-CoV-2 coronavirus pandemic in China in late 2019 led to the fast development of efficient therapeutics. Of the major structural proteins encoded by the SARS-CoV-2 genome, the SPIKE (S) protein has attracted considerable research interest because of the central role it plays in virus entry into host cells. Therefore, to date, most immunization strategies aim at inducing neutralizing antibodies against the surface viral S protein. The SARS-CoV-2 S protein is heavily glycosylated with 22 predicted N-glycosylation consensus sites as well as numerous mucin-type O-glycosylation sites. As a consequence, O- and N-glycosylations of this viral protein have received particular attention. Glycans N-linked to the S protein are mainly exposed at the surface and form a shield-masking specific epitope to escape the virus antigenic recognition. In this work, the N-glycosylation status of the S protein within virus-like particles (VLPs) produced in Nicotiana benthamiana (N. benthamiana) was investigated using a glycoproteomic approach. We show that 20 among the 22 predicted N-glycosylation sites are dominated by complex plant N-glycans and one carries oligomannoses. This suggests that the SARS-CoV-2 S protein produced in N. benthamiana adopts an overall 3D structure similar to that of recombinant homologues produced in mammalian cells.

show abstract

“…As in mammals, plants can also produce proteins displaying O-glycosylation profiles (Tekoah, 2006). However, the O-glycosylation profiles of the recombinant proteins produced from plant-based expression systems were only poorly described in the literature (Karnoup et al, 2005;Kim et al, 2016;Schoberer and Strasser, 2017). O-linked glycosylation can be found in amino acids that contain a hydroxyl group (i.e.…”

Section: Discussionmentioning

confidence: 99%

“…Like N-linked glycans, O-linked glycans play an important role in protein stability and function (Kim et al, 2016;Zhang et al, 2013). In order to further characterize the glycosylation profile of the rIDUA_RLT protein produced, an analysis of its O-glycosylation profile was performed.…”

Section: O-glycosylation Analysis Of Ridua_rlt Produced In the Hairy mentioning

confidence: 99%

Brassica rapa hairy root based expression system leads to the production of highly homogenous and reproducible profiles of recombinant human alpha‐L‐iduronidase

Cardon¹,

Pallisse²,

Bardor

et al. 2018

Plant Biotechnology Journal

View full text Add to dashboard Cite

The Brassica rapa hairy root based expression platform, a turnip hairy root based expression system, is able to produce human complex glycoproteins such as the alpha-L-iduronidase (IDUA) with an activity similar to the one produced by Chinese Hamster Ovary (CHO) cells. In this article, a particular attention has been paid to the N- and O-glycosylation that characterize the alpha-L-iduronidase produced using this hairy root based system. This analysis showed that the recombinant protein is characterized by highly homogeneous post translational profiles enabling a strong batch to batch reproducibility. Indeed, on each of the 6 N-glycosylation sites of the IDUA, a single N-glycan composed of a core Man GlcNAc carrying one beta(1,2)-xylose and one alpha(1,3)-fucose epitope (M3XFGN2) was identified, highlighting the high homogeneity of the production system. Hydroxylation of proline residues and arabinosylation were identified during O-glycosylation analysis, still with a remarkable reproducibility. This platform is thus positioned as an effective and consistent expression system for the production of human complex therapeutic proteins.

show abstract

O-glycans and O-glycosylation sites of recombinant human GM-CSF derived from suspension-cultured rice cells, and their structural role

Cited by 10 publications

References 35 publications

An etanercept O-glycovariant with enhanced potency

An etanercept O-glycovariant with enhanced potency

Investigation of the N-Glycosylation of the SARS-CoV-2 S Protein Contained in VLPs Produced in Nicotiana benthamiana

Brassica rapa hairy root based expression system leads to the production of highly homogenous and reproducible profiles of recombinant human alpha‐L‐iduronidase

Contact Info

Product

Resources

About