Work

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

Year: 2024

Type: article

Abstract: 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 • Detailed protocol for culturing primary cells of intrahepatic cholangiocarcinoma • Sorting cancer stem cells from intrahepatic cholangiocarcinoma tumor samples • α-1,2-Mannosylated CD133 is a marker for sorting and identifying cancer stem cells Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. 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.

Source: STAR Protocols

DOI: 10.1016/j.xpro.2024.103443