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Fragmentation stability and retention time-shift obtained by LC-MS/MS to distinguish sialylated N-glycan linkage isomers in therapeutic glycoproteins

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Journal of Pharmaceutical Analysis 2023年第3期13卷 305-314页

作者： Chi Soo Park Minju Kang Ahyeon Kim chulmin moon Mirae Kim Jieun Kim Subin Yang Leeseul Jang Ji Yeon Jang Ha Hyung Kim Department of Global Innovative Drugs Graduate School of Chung-Ang UniversitySeoul06974Republic of Korea

Sialylated N-glycan isomers withα2-3 orα2-6 linkage(s)have distinctive roles in glycoproteins,but are difficult to ***-type(WT)and glycoengineered(mutant)therapeutic glycoproteins,cytotoxic T lymphocyte-associated antigen-4-immunoglobulin(CTLA4-Ig),were produced in Chinese hamster ovary cell lines;however,their linkage isomers have not been *** this study,N-glycans of CTLA4-Igs were released,labeled with procainamide,and analyzed by liquid chromatography-tandem mass spectrometry(MS/MS)to identify and quantify sialylated N-glycan linkage *** linkage isomers were distinguished by comparison of 1)intensity of the N-acetylglucosamine ion to the sialic acid ion(Ln/Nn)using different fragmentation stability in MS/MS spectra and 2)retention time-shift for a selective m/z value in the extracted ion *** isomer was distinctively identified,and each quantity(>0.1%)was obtained relative to the total N-glycans(100%)for all observed ionization *** sialylated N-glycan isomers with onlyα2-3 linkage(s)in WT were identified,and each isomer's sum of quantities was 50.4%.Furthermore,39 sialylated N-glycan isomers(58.8%)in mono-(3 N-glycans;0.9%),bi-(18;48.3%),tri-(14;8.9%),and tetra-(4;0.7%)antennary structures of mutant were obtained,which comprised mono-(15 N-glycans;25.4%),di-(15;28.4%),tri-(8;4.8%),and tetra-(1;0.2%)sialylation,respectively,with onlyα2-3(10 N-glycans;4.8%),bothα2-3 andα2-6(14;18.4%),and onlyα2-6(15;35.6%)linkage(s).These results are consistent with those forα2-3 neuraminidase-treated *** study generated a novel plot of Ln/Nn versus retention time to distinguish sialylated N-glycan linkage isomers in glycoprotein.

关键词： Therapeutic glycoprotein Sialylation Linkage isomer LC-MS/MS

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Polarization-sensitive intensity diffraction tomography

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Light(Science & Applications) 2023年第6期12卷 1071-1083页

作者： Seungri Song Jeongsoo Kim Taegyun moon Baekcheon Seong Woovin Kim Chang-Hyuk Yoo Jun-Kyu Choi chulmin Joo Department of Mechanical Engineering Yonsei UniversitySeoul 03722Republic of Korea Small Machines Company Ltd.Seoul 04808Republic of Korea

Optical anisotropy,which is an intrinsic property of many materials,originates from the structural arrangement of molecular structures,and to date,various polarization-sensitive imaging(PSI)methods have been developed to investigate the nature of anisotropic *** particular,the recently developed tomographic PSI technologies enable the investigation of anisotropic materials through volumetric mappings of the anisotropy distribution of these ***,these reported methods mostly operate on a single scattering model,and are thus not suitable for three-dimensional(3D)PSI imaging of multiple scattering ***,we present a novel reference-free 3D polarization-sensitive computational imaging technique—polarization-sensitive intensity diffraction tomography(PS-IDT)—that enables the reconstruction of 3D anisotropy distribution of both weakly and multiple scattering specimens from multiple intensity-only measurements.A 3D anisotropic object is illuminated by circularly polarized plane waves at various illumination angles to encode the isotropic and anisotropic structural information into 2D intensity *** information are then recorded separately through two orthogonal analyzer states,and a 3D Jones matrix is iteratively reconstructed based on the vectorial multi-slice beam propagation model and gradient descent *** demonstrate the 3D anisotropy imaging capabilities of PS-IDT by presenting 3D anisotropy maps of various samples,including potato starch granules and tardigrade.

关键词： materials anisotropy intensity

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