Antibody variable region glycosylation: biochemical and clinical effects

Wright, Ann; Morrison, Sherie L.

doi:10.1007/bf00201106

Cited by 27 publications

(12 citation statements)

References 56 publications

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“…While antibodies are always glycosylated in the C H 2 domain of Fc, glycosylation can also be observed when complementarity determining regions in the variable domains contain a consensus N-glycosylation sequence motif (25,26). Our initial results prompted further investigation, as potential glycosylation of natural C H 1 constant domain has never been reported, which is in agreement with the fact that no consensus N-linked site is present on this conserved sequence.…”

Section: From the Department Of Process And Product Development Amgesupporting

confidence: 76%

Asparagine-linked Oligosaccharides Present on a Non-consensus Amino Acid Sequence in the CH1 Domain of Human Antibodies

Valliere-Douglass

Kodama

Mujacic

et al. 2009

Journal of Biological Chemistry

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We report that N-linked oligosaccharide structures can be present on an asparagine residue not adhering to the consensus site motif NX(S/T), where X is not proline, described in the literature. We have observed oligosaccharides on a non-consensus asparaginyl residue in the C H 1 constant domain of IgG1 and IgG2 antibodies. The initial findings were obtained from characterization of charge variant populations evident in a recombinant human antibody of the IgG2 subclass. HPLC-MS results indicated that cation-exchange chromatography acidic variant populations were enriched in antibody with a second glycosylation site, in addition to the well documented canonical glycosylation site located in the C H 2 domain. Subsequent tryptic and chymotryptic peptide map data indicated that the second glycosylation site was associated with the amino acid sequence TVSWN 162 SGAL in the C H 1 domain of the antibody. This highly atypical modification is present at levels of 0.5-2.0% on most of the recombinant antibodies that have been tested and has also been observed in IgG1 antibodies derived from human donors. Site-directed mutagenesis of the C H 1 domain sequence in a recombinanthuman IgG1 antibody resulted in an increase in non-consensus glycosylation to 3.15%, a greater than 4-fold increase over the level observed in the wild type, by changing the ؊1 and ؉1 amino acids relative to the asparagine residue at position 162. We believe that further understanding of the phenomenon of non-consensus glycosylation can be used to gain fundamental insights into the fidelity of the cellular glycosylation machinery.

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Section: From the Department Of Process And Product Development Amgesupporting

confidence: 76%

Asparagine-linked Oligosaccharides Present on a Non-consensus Amino Acid Sequence in the CH1 Domain of Human Antibodies

Valliere-Douglass

Kodama

Mujacic

et al. 2009

Journal of Biological Chemistry

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“…Recently, de-fucosylation on the N297-linked glycan in the Fc part of the Ab has been shown to increase ADCC activity, indicating the importance of glyco-engineering of Ab for improved clinical efficacy [23]. On the other hand, Fab N-linked glycosylation in the hypervariable regions, while occurring much less frequently, has been reported to influence the binding affinity of antigens [24][25][26] and may also be involved in IgG self-association, aggregation, and cryo-precipitation [27].…”

Section: Introductionmentioning

confidence: 99%

Endoglycosidase treatment abrogates IgG arthritogenicity: Importance of IgG glycosylation in arthritis

et al. 2007

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The glycosylation status of IgG has been implicated in the pathology of rheumatoid arthritis. Earlier, we reported the identification of a novel secreted endo-b-Nacetylglucosaminidase (EndoS), secreted by Streptococcus pyogenes that specifically hydrolyzes the b-1,4-di-N-acetylchitobiose core of the asparagine-linked glycan of human IgG. Here, we analyzed the arthritogenicity of EndoS-treated collagen type II (CII)-specific mouse mAb in vivo. Endoglycosidase treatment of the antibodies inhibited the induction of arthritis in (BALB/c Â B10.Q) F1 mice and induced a milder arthritis in B10.RIII mice as compared with the severe arthritis induced by non-treated antibodies. Furthermore, EndoS treatment did not affect the binding of IgG to CII and their ability to activate complement, but it resulted in reduced IgG binding to FccR and disturbed the formation of stable immune complexes. Hence, the asparagine-linked glycan on IgG plays a crucial role in the development of arthritis. IntroductionThe impact of glycosylation, one of the most important post-translational modifications, on the structure and biological properties of glycoproteins has been well documented [1,2]. IgG molecules are mainly glycosylated at Asn-297 of the CH2 domain within the Fc region [3,4], with variable galactosylation but limited sialylation. The remaining glycosylation occurs in the hypervariable regions of the Fab region, with the position and frequency of occurrence being dependent on the presence of the consensus sequence Asn-Xaa-Thr/ Ser for N-glycosylation, and is characterized by a high incidence of sialylated structures. Murine IgG contain 2.3 asparagine-linked (N-linked) biantennary oligosaccharide chains per molecule [5], and human IgG contain 2.8 [6]. The minimal oligosaccharide structure is a hexasaccharide (GlcNAc2Man3GlcNAc) with variable sugar residues attached, which results in the generation of the multiple glycoforms. About 30 variants of biantennary chains occur, resulting in Clinical immunology

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“…The Fc glycans of IgG are important for the antibody to maintain its structure and functions (5). In the hypervariable regions, Fab N-linked glycosylation has been reported to influence the binding affinity of antigens (6). Reports have shown that IgG forms containing Fab oligosaccharides may have preferential roles in IgG self-association, aggregation, and cryoprecipitation (7).…”

mentioning

confidence: 98%

A Study of Immunoglobulin G Glycosylation in Monoclonal and Polyclonal Species by Electrospray and Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry

Saba

Kunkel

Jan

et al. 2002

Analytical Biochemistry

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Antibody variable region glycosylation: biochemical and clinical effects

Cited by 27 publications

References 56 publications

Asparagine-linked Oligosaccharides Present on a Non-consensus Amino Acid Sequence in the CH1 Domain of Human Antibodies

Asparagine-linked Oligosaccharides Present on a Non-consensus Amino Acid Sequence in the CH1 Domain of Human Antibodies

Endoglycosidase treatment abrogates IgG arthritogenicity: Importance of IgG glycosylation in arthritis

A Study of Immunoglobulin G Glycosylation in Monoclonal and Polyclonal Species by Electrospray and Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry

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