Impact of N-glycosylation on Fcγ Receptor / IgG Interactions: Unravelling Differences with an Enhanced Surface Plasmon Resonance Biosensor Assay Based on Coiled-coil Interactions

Overview

Journal MAbs

Specialty Allergy & Immunology

Date 2019 Mar 2

PMID 30822189

Citations 9

Authors

Florian Cambay

Olivier Henry

Yves Durocher

Gregory De Crescenzo

Affiliations

Soon will be listed here.

Abstract

The N-glycosylation profile of immunoglobulin G (IgG) is considered a critical quality attribute due to its impact on IgG-Fc gamma receptor (FcγR) interactions, which subsequently affect antibody-dependent cell-based immune responses. In this study, we investigated the impact of the FcγR capture method, as well as FcγR N-glycosylation, on the kinetics of interaction with various glycoforms of trastuzumab (TZM) in a surface plasmon resonance (SPR) biosensor assay. More specifically, we developed a novel strategy based on coiled-coil interactions for the stable and oriented capture of coil-tagged FcγRs at the biosensor surface. Coil-tagged FcγR capture outperformed all other capture strategies applied to the SPR study of IgG-FcγR interactions, as the robustness and reproducibility of the assay and the shelf life of the biosensor chip were excellent (> 1,000 IgG injections with the same biosensor surface). Coil-tagged FcγRs displaying different N-glycosylation profiles were generated either by different expression systems, in vitro glycoengineering or by size-exclusion chromatography, and roughly characterized by lectin blotting. Of salient interest, the overlay of their kinetics of interaction with several TZM glycoforms revealed key differences on both association and dissociation kinetics, confirming a complex influence of the FcγR N-glycosylation and its inherent heterogeneity upon receptor interaction with mAbs. This work is thus an important step towards better understanding of the impact of glycosylation upon binding of IgGs, either natural or engineered, to their receptors.

Citing Articles

A Biosensor Assay Based on Coiled-Coil-Mediated Human ACE2 Receptor Capture for the Analysis of Its Interactions with the SARS-CoV-2 Receptor Binding Domain.

Forest-Nault C, Koyuturk I, Gaudreault J, Pelletier A, LAbbe D, Cass B Methods Mol Biol. 2024; 2762:89-105.

PMID: 38315361 DOI: 10.1007/978-1-0716-3666-4_6.

Multi-temperature experiments to ease analysis of heterogeneous binder solutions by surface plasmon resonance biosensing.

Gaudreault J, Durocher Y, Henry O, De Crescenzo G Sci Rep. 2022; 12(1):14401.

PMID: 36002549 PMC: 9402583. DOI: 10.1038/s41598-022-18450-y.

Impact of the temperature on the interactions between common variants of the SARS-CoV-2 receptor binding domain and the human ACE2.

Forest-Nault C, Koyuturk I, Gaudreault J, Pelletier A, LAbbe D, Cass B Sci Rep. 2022; 12(1):11520.

PMID: 35798770 PMC: 9261887. DOI: 10.1038/s41598-022-15215-5.

Impact of IgG1 N-glycosylation on their interaction with Fc gamma receptors.

Cambay F, Raymond C, Brochu D, Gilbert M, Tu T, Cantin C Curr Res Immunol. 2022; 1:23-37.

PMID: 35493857 PMC: 9040152. DOI: 10.1016/j.crimmu.2020.06.001.

Quantitative N-glycoproteome analysis of bovine milk and yogurt.

Xiao J, Wang J, Gan R, Wu D, Xu Y, Peng L Curr Res Food Sci. 2022; 5:182-190.

PMID: 35072106 PMC: 8763629. DOI: 10.1016/j.crfs.2022.01.003.

References

Durocher Y, Perret S, Kamen A . High-level and high-throughput recombinant protein production by transient transfection of suspension-growing human 293-EBNA1 cells. Nucleic Acids Res. 2002; 30(2):E9. PMC: 99848. DOI: 10.1093/nar/30.2.e9. View

Radaev S, Sun P . Recognition of immunoglobulins by Fcgamma receptors. Mol Immunol. 2002; 38(14):1073-83. DOI: 10.1016/s0161-5890(02)00036-6. View

Litowski J, Hodges R . Designing heterodimeric two-stranded alpha-helical coiled-coils: the effect of chain length on protein folding, stability and specificity. J Pept Res. 2002; 58(6):477-92. DOI: 10.1034/j.1399-3011.2001.10972.x. View

Takahashi N, Cohen-Solal J, Galinha A, Fridman W, Sautes-Fridman C, Kato K . N-glycosylation profile of recombinant human soluble Fcgamma receptor III. Glycobiology. 2002; 12(8):507-15. DOI: 10.1093/glycob/cwf063. View

De Crescenzo G, Litowski J, Hodges R, OConnor-McCourt M . Real-time monitoring of the interactions of two-stranded de novo designed coiled-coils: effect of chain length on the kinetic and thermodynamic constants of binding. Biochemistry. 2003; 42(6):1754-63. DOI: 10.1021/bi0268450. View

De Crescenzo G, Pham P, Durocher Y, OConnor-McCourt M . Transforming growth factor-beta (TGF-beta) binding to the extracellular domain of the type II TGF-beta receptor: receptor capture on a biosensor surface using a new coiled-coil capture system demonstrates that avidity contributes significantly to.... J Mol Biol. 2003; 328(5):1173-83. DOI: 10.1016/s0022-2836(03)00360-7. View

Okazaki A, Shoji-Hosaka E, Nakamura K, Wakitani M, Uchida K, Kakita S . Fucose depletion from human IgG1 oligosaccharide enhances binding enthalpy and association rate between IgG1 and FcgammaRIIIa. J Mol Biol. 2004; 336(5):1239-49. DOI: 10.1016/j.jmb.2004.01.007. View

De Crescenzo G, Pham P, Durocher Y, Chao H, OConnor-McCourt M . Enhancement of the antagonistic potency of transforming growth factor-beta receptor extracellular domains by coiled coil-induced homo- and heterodimerization. J Biol Chem. 2004; 279(25):26013-8. DOI: 10.1074/jbc.M400655200. View

Yamane-Ohnuki N, Kinoshita S, Inoue-Urakubo M, Kusunoki M, Iida S, Nakano R . Establishment of FUT8 knockout Chinese hamster ovary cells: an ideal host cell line for producing completely defucosylated antibodies with enhanced antibody-dependent cellular cytotoxicity. Biotechnol Bioeng. 2004; 87(5):614-22. DOI: 10.1002/bit.20151. View

10.

Warnock D, Bai X, Autote K, Gonzales J, Kinealy K, Yan B . In vitro galactosylation of human IgG at 1 kg scale using recombinant galactosyltransferase. Biotechnol Bioeng. 2005; 92(7):831-42. DOI: 10.1002/bit.20658. View

11.

Ferrara C, Stuart F, Sondermann P, Brunker P, Umana P . The carbohydrate at FcgammaRIIIa Asn-162. An element required for high affinity binding to non-fucosylated IgG glycoforms. J Biol Chem. 2005; 281(8):5032-6. DOI: 10.1074/jbc.M510171200. View

12.

Kanda Y, Yamada T, Mori K, Okazaki A, Inoue M, Kitajima-Miyama K . Comparison of biological activity among nonfucosylated therapeutic IgG1 antibodies with three different N-linked Fc oligosaccharides: the high-mannose, hybrid, and complex types. Glycobiology. 2006; 17(1):104-18. DOI: 10.1093/glycob/cwl057. View

13.

Li P, Jiang N, Nagarajan S, Wohlhueter R, Selvaraj P, Zhu C . Affinity and kinetic analysis of Fcgamma receptor IIIa (CD16a) binding to IgG ligands. J Biol Chem. 2007; 282(9):6210-21. DOI: 10.1074/jbc.M609064200. View

14.

Jefferis R . Antibody therapeutics: isotype and glycoform selection. Expert Opin Biol Ther. 2007; 7(9):1401-13. DOI: 10.1517/14712598.7.9.1401. View

15.

Boucher C, St-Laurent G, Loignon M, Jolicoeur M, De Crescenzo G, Durocher Y . The bioactivity and receptor affinity of recombinant tagged EGF designed for tissue engineering applications is defined by the nature and position of the tags. Tissue Eng Part A. 2008; 14(12):2069-77. DOI: 10.1089/ten.tea.2008.0037. View

16.

Shibata-Koyama M, Iida S, Okazaki A, Mori K, Kitajima-Miyama K, Saitou S . The N-linked oligosaccharide at Fc gamma RIIIa Asn-45: an inhibitory element for high Fc gamma RIIIa binding affinity to IgG glycoforms lacking core fucosylation. Glycobiology. 2008; 19(2):126-34. PMC: 2639328. DOI: 10.1093/glycob/cwn110. View

17.

Zhang J, Liu X, Bell A, To R, Baral T, Azizi A . Transient expression and purification of chimeric heavy chain antibodies. Protein Expr Purif. 2008; 65(1):77-82. DOI: 10.1016/j.pep.2008.10.011. View

18.

Bruhns P, Iannascoli B, England P, Mancardi D, Fernandez N, Jorieux S . Specificity and affinity of human Fcgamma receptors and their polymorphic variants for human IgG subclasses. Blood. 2008; 113(16):3716-25. DOI: 10.1182/blood-2008-09-179754. View

19.

Shibata-Koyama M, Iida S, Misaka H, Mori K, Yano K, Shitara K . Nonfucosylated rituximab potentiates human neutrophil phagocytosis through its high binding for FcgammaRIIIb and MHC class II expression on the phagocytotic neutrophils. Exp Hematol. 2009; 37(3):309-21. DOI: 10.1016/j.exphem.2008.11.006. View

20.

Boucher C, St-Laurent G, Jolicoeur M, De Crescenzo G, Durocher Y . Protein detection by Western blot via coiled-coil interactions. Anal Biochem. 2009; 399(1):138-40. DOI: 10.1016/j.ab.2009.12.007. View