Protocol for assessing murine cell doublet engagement and subsequent effects using flow cytometry and imaging flow cytometry
Year: 2024
Type: article
Abstract: Physical interactions between two immune cells or between immune and cancer cells play a major role in shaping the immune response in the tumor microenvironment, making them prime therapeutic targets for bispecific engagers. Here, we present a protocol for assessing murine cell doublet engagement and subsequent effects using flow cytometry and imaging flow cytometry. We describe steps for identifying bispecific cell engager antibodies at the cell-cell interface, doublet quantification, and characterizing cellular protein morphology and processes within the doublet. For complete details on the use and execution of this protocol, please refer to Shapir Itai et al. 1 • Two approaches for determining physical interactions between two cells • In vitro system and flow cytometry protocol for cell binding analysis • Imaging flow cytometry protocol for engager localization at the cell-cell interface • Applications to assess cell activation, internalization, synapse formation, or death Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. Physical interactions between two immune cells or between immune and cancer cells play a major role in shaping the immune response in the tumor microenvironment, making them prime therapeutic targets for bispecific engagers. Here, we present a protocol for assessing murine cell doublet engagement and subsequent effects using flow cytometry and imaging flow cytometry. We describe steps for identifying bispecific cell engager antibodies at the cell-cell interface, doublet quantification, and characterizing cellular protein morphology and processes within the doublet.
Source: STAR Protocols
Authors: Yuval Shapir Itai, Ziv Porat, Rony Dahan
Institution: Weizmann Institute of Science
Language: English
Subfield: Radiology, Nuclear Medicine and Imaging
Field: Medicine
Domain: Health Sciences
Open Access status: gold
Estimated APC paid: $1,500
OpenAlex ID: W4402679795