Work

Protocol to identify and sort antigen-specific B cells from Plasmodium-infected mice

Year: 2025

Type: article

Abstract: Here, we detail a protocol to identify an antigen-specific B cell population using SpyCage reagents tailored to bind MSP 1-19 antigen in a rodent malaria model. We describe steps for the preparation of splenic single-cell suspension, B cell enrichment, and staining to facilitate sorting of live cells for downstream applications such as single-cell RNA, V(D)J, and assay for transposase-accessible chromatin (ATAC) sequencing. For complete details on the use and execution of this protocol, please refer to Calôba et al. 1 • Steps for preparing single-cell suspension and antigen-specific B cell enrichment • Instructions for optimizing staining of compensation/reference control beads • Guide to sort-purify antigen-specific B cells for downstream workflow Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. Here, we detail a protocol to identify an antigen-specific B cell population using SpyCage reagents tailored to bind MSP 1-19 antigen in a rodent malaria model. We describe steps for the preparation of splenic single-cell suspension, B cell enrichment, and staining to facilitate sorting of live cells for downstream applications such as single-cell RNA, V(D)J, and assay for transposase-accessible chromatin (ATAC) sequencing.

Source: STAR Protocols

DOI: 10.1016/j.xpro.2025.104129

Authors: Carolina Calôba, Allen M. Minns, Scott E. Lindner, Rahul Vijay

Institutions: Rosalind Franklin University of Medicine and Science, Pennsylvania State University

Language: English

Topic: Malaria Research and Control

Subfield: Public Health, Environmental and Occupational Health

Field: Medicine

Domain: Health Sciences

SDG: Good health and well-being

Open Access status: gold

Estimated APC paid: $1,550