Cell surface profiling of cultured cells by direct hydrazide capture of oxidized glycoproteins.
MethodsX
; 11: 102349, 2023 Dec.
Article
em En
| MEDLINE
| ID: mdl-37680365
ABSTRACT
Glycoproteins are a particularly interesting subset of the cellular proteome as a high proportion of proteins present on the extracellular cell surface are glycosylated. These cell surface proteins are ideal targets for biologic drug therapies or for diagnostics tests. Here, we describe a modification of the well-described Cell Surface Capture (CSC) method for the selective isolation and identification of cell surface glycoproteins that contain N-linked carbohydrates. This modification, which we refer to as Direct Cell Surface Capture (D-CSC), is based on oxidation of cell surface glycans on intact cells, followed by direct conjugation of the oxidized oligosaccharides to a solid support using hydrazide chemistry, with no biotinylation step. As a proof-of-principle, we applied D-CSC to the analysis of cell surface membrane proteins of three adherent cancer cell lines (A549, OVCAR3, and U87MG) and compared our results to those published using the well-established Cell Surface Capture (CSC) method, demonstrating comparable selectivity for cell surface proteins. â¢A method enabling the identification of cell surface proteins from cells in culture is described.â¢Application of this method to profile the cell surface on three different cancer cell lines is included.
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Idioma:
En
Revista:
MethodsX
Ano de publicação:
2023
Tipo de documento:
Article
País de afiliação:
Canadá