Protocol for sorting cancer stem cells using a combination of anti-CD133 antibody and lectin cyanovirin-N

Yuanyan Wei; Shuting Geng; Qihang Chen; Jiayue Chen; Zhijun Fan; Yi Li; Yu Si; Yuerong Yang; Ziwei Liang; Jianhai Jiang

STAR Protocols (Dec 2024)

Protocol for sorting cancer stem cells using a combination of anti-CD133 antibody and lectin cyanovirin-N

Yuanyan Wei,
Shuting Geng,
Qihang Chen,
Jiayue Chen,
Zhijun Fan,
Yi Li,
Yu Si,
Yuerong Yang,
Ziwei Liang,
Jianhai Jiang

Affiliations

Yuanyan Wei: NHC Key Laboratory of Glycoconjuates Research, Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Fudan University, Shanghai 200032, P.R. China; Corresponding author
Shuting Geng: NHC Key Laboratory of Glycoconjuates Research, Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Fudan University, Shanghai 200032, P.R. China
Qihang Chen: NHC Key Laboratory of Glycoconjuates Research, Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Fudan University, Shanghai 200032, P.R. China
Jiayue Chen: NHC Key Laboratory of Glycoconjuates Research, Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Fudan University, Shanghai 200032, P.R. China
Zhijun Fan: NHC Key Laboratory of Glycoconjuates Research, Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Fudan University, Shanghai 200032, P.R. China
Yi Li: NHC Key Laboratory of Glycoconjuates Research, Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Fudan University, Shanghai 200032, P.R. China
Yu Si: NHC Key Laboratory of Glycoconjuates Research, Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Fudan University, Shanghai 200032, P.R. China
Yuerong Yang: NHC Key Laboratory of Glycoconjuates Research, Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Fudan University, Shanghai 200032, P.R. China
Ziwei Liang: NHC Key Laboratory of Glycoconjuates Research, Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Fudan University, Shanghai 200032, P.R. China; Department of Biomedical Engineering, Research Center for Nano-biomaterials & Regenerative Medicine, College of Artificial Intelligence, Shanxi Key Laboratory of Materials Strength & Structural Impact, Taiyuan University of Technology, Taiyuan 030024, China
Jianhai Jiang: NHC Key Laboratory of Glycoconjuates Research, Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Fudan University, Shanghai 200032, P.R. China; Corresponding author

Journal volume & issue: Vol. 5, no. 4
p. 103443

Abstract

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Summary: CD133 is widely used as a marker to isolate cancer stem cells (CSCs). However, the structural ambiguity of N-glycan of CD133 limits its application in the isolation of CSCs. Here, we present a protocol to sort CSCs from tumor samples by combining CD133 with α-1,2-high-mannose type glycan chains. We describe steps for purifying and biotinylating cyanovirin-N (CVN) proteins and sorting tumor cells and CD133+α-1,2-Man+ (α-1,2-linked mannose+) cells to provide more specific markers for CSCs.For complete details on the use and execution of this protocol, please refer to Wei et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

Published in STAR Protocols

ISSN: 2666-1667 (Online)
Publisher: Elsevier
Country of publisher: United States
LCC subjects: Science: Science (General)
Website: https://star-protocols.cell.com/

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