Development of Immobilized Enzyme Reactors for the characterization of the glycosylation heterogeneity of a protein

Stan Perchepied , Nicolas Eskenazi (1) , Chiara Giangrande (1) , Julien Camperi (2) , Thierry Fournier , Joelle Vinh (1) , Nathalie N. Delaunay (3) , Valérie Pichon (4)

1 SMBP - Spectrométrie de Masse Biologique et Protéomique (USR3149 / FRE2032)
2 LSABM - Laboratoire Sciences Analytiques, Bioanalytiques, et Miniaturisation
3 CBI - Chimie-Biologie-Innovation (UMR 8231)
4 UPMC - Université Pierre et Marie Curie - Paris 6

Abbreviations: ACN, acetonitrile; BCA, bicinchoninic acid assay; BPC, base peak chromatogram; DTT, dithiothreitol; E/S, enzyme to substrate ratio; EIC, extracted ion chromatogram; FA, formic acid; Fuc, fucose; GalNAc, N-acetylgalactosamine; hCG, human chorionic gonadotropin; hCGα, alpha subunit of the human chorionic gonadotropin; hCGβ, beta subunit of the human chorionic gonadotropin; Hex, hexose; HexNAc, N-acetylhexoseamine; IAM, iodoacetamide; IMER, Immobilized Enzyme Reactor; NeuAc, N-acetylneuraminic acid; NeuGc, N-glycolylneuraminic acid; MS/MS, tandem mass spectrometry; P-IMER, pepsin-based immobilized enzyme reactor; PTM, post-translational modification; r-hCG, recombinant human chorionic gonadotropin; TFA, trifluoroacetic acid; TIMER , trypsin-based immobilized enzyme reactor; TRIS, Trizma hydrochloride; u-hCG, human chorionic gonadotropin isolated from urine of pregnant women

Mots clés: Immobilized Enzyme Reactor; Glycoproteomics; Glycosylation mapping; Human chorionic gonadotropin; NanoLC-MS/MS

Domaines: Chimie; Chimie analytique

Stan Perchepied, Nicolas Eskenazi, Chiara Giangrande, Julien Camperi, Thierry Fournier, et al.. Development of Immobilized Enzyme Reactors for the characterization of the glycosylation heterogeneity of a protein. Talanta, 2020, 206, pp.120171. ⟨10.1016/j.talanta.2019.120171⟩. ⟨hal-02330897⟩