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

## TL;DR

This paper describes a detailed protocol for isolating B cells specific to a malaria antigen in mice for advanced genetic and chromatin analysis.

## Contribution

The protocol introduces a method using SpyCage reagents to target the MSP1-19 antigen in rodent malaria models for downstream single-cell sequencing.

## Key findings

- Steps for preparing single-cell suspensions and enriching antigen-specific B cells are outlined.
- Optimization of staining and sorting procedures is detailed for downstream RNA and ATAC sequencing.
- The protocol facilitates isolation of live B cells for multi-omics analysis.

## Abstract

Here, we detail a protocol to identify an antigen-specific B cell population using SpyCage reagents tailored to bind MSP1-19 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

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 MSP1-19 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.