Earticle

초록

영어

Agalsidase beta, recombinant enzyme therapeutic, which is a glycoprotein that treats lysosome accumulation disorders caused by deficiency of the enzyme alpha galactosidase A has 3 glycosylation sites(N108, N161, N184). In this protein, there are various glycans from acidic glycans to high mannose, so accurate monitoring is required for proper afficacy and function of drug. Recently, intact analysis is heavily used due to the advantage of simple preparation, short MS analysis time, glycan occupancy and micro-heterogeneity confirmation. However, numerous glycosylation sites, high molecular weight and glycan heterogeneity complicate the mass spectrum, making data interpretation and glycan assignment difficult. Here, we obtained the mass value of intact agalsidase beta, which eased the complexity of the mass spectrum by lowering the high molecular weight of the glycoprotein through stepwise enzymatic treatment. To facilitate the assignment of sialic acid and phosphorylated glycans at each glycosylation sites, deconvoluted spectra of sialidase and phosphatase-treated and untreated agalsidase beta were compared. Possible glycoforms could be assigned for each peak in the deconvoluted spectra within the accepted mass tolerance of ±5 Da. In the monomer and dimer structures of agalsidase beta, a glycosylation pattern was confirmed in which HexNAcNeuAc residues(494.1748 Da) and 2HexNAc2NeuAc residues (988.3496 Da) were increased, respectively. Our approach can be a efficient tool to confirm glycosylation pattern of therapeutic glycoproteins in biosimilar development.

저자

• GaHyeon Kim [ Graduate School of Analytical Science and Technology, Chungnam National University, Asia-Pacific Glycomics Reference Site, Daejeon, 34134, Korea ]
• Jung A Seo [ Graduate School of Analytical Science and Technology, Chungnam National University, Asia-Pacific Glycomics Reference Site, Daejeon, 34134, Korea ]
• Myung Jin Oh [ Graduate School of Analytical Science and Technology, Chungnam National University, Asia-Pacific Glycomics Reference Site, Daejeon, 34134, Korea ]
• Hyun Joo An [ Graduate School of Analytical Science and Technology, Chungnam National University, Asia-Pacific Glycomics Reference Site, Daejeon, 34134, Korea ]

참고문헌

발행기관

간행물

표지보기 관심저널 등록

• 간행물명

  한국당과학회 학술대회

• 간기

  연간

• 수록기간

  2006~2022

• 십진분류

  KDC 517 DDC 614