公开(公告)号：US20180106780A1

公开(公告)日：2018-04-19

申请号：US15729317

申请日：2017-10-10

Applicant: Academia Sinica

Inventor： Chi-Huey WONG ,  Tsui-Ling HSU ,  Sarah R. Hanson ,  Masaaki SAWA

IPC: G01N33/50 ,  G01N33/533

CPC classification number: G01N33/5005 ,  G01N33/5008 ,  G01N33/533 ,  G01N2400/00

Abstract: Methods for metabolic oligosaccharide engineering that incorporates derivatized alkyne-bearing sugar analogs as “tags” into cellular glycoconjugates are disclosed. Alkynyl derivatized Fuc and alkynyl derivatized ManNAc sugars are incorporated into cellular glycoconjugates. Chemical probes comprising an azide group and a visual or fluorogenic probe and used to label alkyne-derivatized sugar-tagged glycoconjugates are disclosed. Chemical probes bind covalently to the alkynyl group by Cu(I)-catalyzed [3+2] azide-alkyne cycloaddition and are visualized at the cell surface, intracellularly, or in a cellular extract. The labeled glycoconjugate is capable of detection by flow cytometry, SDS-PAGE, Western blot, ELISA, confocal microscopy, and mass spectrometry.

公开(公告)号：US20190269227A1

公开(公告)日：2019-09-05

申请号：US16415411

申请日：2019-05-17

Applicant: ACADEMIA SINICA

Inventor： Chi-Huey WONG ,  Tsui-Ling HSU ,  Sarah R. HANSON ,  Masaaki SAWA

IPC: A45F3/14 ,  A41F9/02 ,  A45F5/02 ,  A45F3/00 ,  A45F3/10

Abstract: The present disclosure relates to a method for metabolic oligosaccharide engineering that incorporates derivatized alkyne-bearing sugar analogs as “tags” into cellular glycoconjugates. The disclosed method incorporates alkynyl derivatized Fuc and alkynyl derivatized ManNAc sugars into a cellular glycoconjugate. A chemical probe comprising an azide group and a visual probe or a fluorogenic probe is used to label the alkyne-derivatized sugar-tagged glycoconjugate. In one aspect, the chemical probe binds covalently to the alkynyl group by Cu(I)-catalyzed [3+2] azide-alkyne cycloaddition and is visualized at the cell surface, intracellularly, or in a cellular extract. The labeled glycoconjugate is capable of detection by flow cytometry, SDS-PAGE, Western blot, ELISA or confocal microscopy, and mass spectrometry.