GlycoVis: Visualizing glycan distribution in the protein <i>N</i>‐glycosylation pathway in mammalian cells

Hossler, Patrick; Goh, Lin Tang; Hu, Wei Shou

doi:10.1002/bit.21062

Cited by 40 publications

(33 citation statements)

References 57 publications

(44 reference statements)

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“…The studies on sialyltransferases' enzymatic activity usually consider G2F or Galb1-4GlcNAc as substrates, and data comparing G1F isomers to G2F as acceptor substrates are missing. 54 Similarly, to our knowledge, G1FS1 has not been studied as acceptor substrate for the galactosyltransferases. 52,[55][56][57] Thus, there is no data in the literature indicating that the presence of a sialic acid on one branch would be an obstacle for galactosylation of the other branch.…”

Section: Discussionmentioning

confidence: 95%

Production of α2,6-sialylated IgG1 in CHO cells

Raymond

Robotham

Spearman

et al. 2015

mAbs

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The presence of a2,6-sialic acids on the Fc N-glycan provides anti-inflammatory properties to the IgGs through a mechanism that remains unclear. Fc-sialylated IgGs are rare in humans as well as in industrial host cell lines such as Chinese hamster ovary (CHO) cells. Facilitated access to well-characterized a2,6-sialylated IgGs would help elucidate the mechanism of this intriguing IgG's effector function. This study presents a method for the efficient Fc glycan a2,6-sialylation of a wild-type and a F243A IgG1 mutant by transient co-expression with the human a2,6-sialyltransferase 1 (ST6) and b1,4-galactosyltransferase 1 (GT) in CHO cells. Overexpression of ST6 alone only had a moderate effect on the glycoprofiles, whereas GT alone greatly enhanced Fc-galactosylation, but not sialylation. Overexpression of both GT and ST6 was necessary to obtain a glycoprofile dominated by a2,6-sialylated glycans in both antibodies. The wild-type was composed of the G2FS(6)1 glycan (38%) with remaining unsialylated glycans, while the mutant glycoprofile was essentially composed of G2FS(6)1 (25%), G2FS(3,6)2 (16%) and G2FS(6,6)2 (37%). The a2,6-linked sialic acids represented over 85% of all sialic acids in both antibodies. We discuss how the limited sialylation level in the wild-type IgG1 expressed alone or with GT results from the glycan interaction with Fc's amino acid residues or from intrinsic galactosyl-and sialyl-transferases substrate specificities.

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Section: Discussionmentioning

confidence: 95%

Production of α2,6-sialylated IgG1 in CHO cells

Raymond

Robotham

Spearman

et al. 2015

mAbs

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“…A simplified numeric representation instead of a text-based representation of glycan structures can further advance the development of computer aided analysis tools to study such a complex network [27]. We have demonstrated that the current numeric format can be easily applied to constructing and visualizing networks of glycan interactions.…”

Section: Discussionmentioning

confidence: 95%

“…Figure 6 exhibits the resulting network, which is a highly branched structure. It should be noted that the current network is an approximation of the glycosylation pathway in CHO cells since the enzymatic requirements and restrictions [27] were not fully considered during the network construction.…”

Section: Constructing Glycosylation Networkmentioning

confidence: 99%

An alpha-numeric code for representing N-linked glycan structures in secreted glycoproteins

Yusufi

Park

Lee

2008

Bioprocess Biosyst Eng

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Advances in high-throughput techniques have led to the creation of increasing amounts of glycome data. The storage and analysis of this data would benefit greatly from a compact notation for describing glycan structures that can be easily stored and interpreted by computers. Towards this end, we propose a fixed-length alpha-numeric code for representing N-linked glycan structures commonly found in secreted glycoproteins from mammalian cell cultures. This code, GlycoDigit, employs a pre-assigned alpha-numeric index to represent the monosaccharides attached in different branches to the core glycan structure. The present branch-centric representation allows us to visualize the structure while the numerical nature of the code makes it machine readable. In addition, a difference operator can be defined to quantitatively differentiate between glycan structures for further analysis. The usefulness and applicability of GlycoDigit were demonstrated by constructing and visualizing an N-linked glycosylation network.

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“…The program also traces all possible reaction paths leading to each glycan and identifies each pathway on the map. Use of the program is illustrated with MALDI-TOF data from permethylated glycans from IgG and tissue plasminogen activator (TPA) (Hossler et al, 2006).…”

Section: Analysis Of Carbohydrates and Glycoconjugatesmentioning

confidence: 99%

Analysis of carbohydrates and glycoconjugates by matrix‐assisted laser desorption/ionization mass spectrometry: An update for the period 2005–2006

Harvey

2010

Mass Spectrometry Reviews

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This review is the fourth update of the original review, published in 1999, on the application of MALDI mass spectrometry to the analysis of carbohydrates and glycoconjugates and brings coverage of the literature to the end of 2006. The review covers fundamental studies, fragmentation of carbohydrate ions, method developments, and applications of the technique to the analysis of different types of carbohydrate. Specific compound classes that are covered include carbohydrate polymers from plants, N- and O-linked glycans from glycoproteins, glycated proteins, glycolipids from bacteria, glycosides, and various other natural products. There is a short section on the use of MALDI-TOF mass spectrometry for the study of enzymes involved in glycan processing, a section on industrial processes, particularly the development of biopharmaceuticals and a section on the use of MALDI-MS to monitor products of chemical synthesis of carbohydrates. Large carbohydrate-protein complexes and glycodendrimers are highlighted in this final section.

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GlycoVis: Visualizing glycan distribution in the protein N‐glycosylation pathway in mammalian cells

Cited by 40 publications

References 57 publications

Production of α2,6-sialylated IgG1 in CHO cells

Production of α2,6-sialylated IgG1 in CHO cells

An alpha-numeric code for representing N-linked glycan structures in secreted glycoproteins

Analysis of carbohydrates and glycoconjugates by matrix‐assisted laser desorption/ionization mass spectrometry: An update for the period 2005–2006

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