Highly efficient and low-cost single-cell culture platform for unbiased analysis of human memory B cell repertoire and antibody discovery
Year: 2025
Type: article
Abstract: The SARS-CoV-2 pandemic underscored the need for innovative approaches to study humoral immunity and isolate monoclonal antibodies (mAbs) with diagnostic and therapeutic potential. Current methods for repertoire analysis at the clonal level require large-scale recombinant mAb production, limiting accessibility and delaying functional insight. We developed a single-cell culture (SCC) platform that enables profiling of human memory B cells and direct recovery of functional mAbs. Using samples from COVID-19 convalescent and vaccinated donors, we optimized SCCs with NB21 feeder cells, R848, and IL-2, achieving efficient clonal expansion and antibody secretion in short-term cultures. Screening and pseudovirus neutralization assays were performed directly with culture supernatants, bypassing the need for early recombinant antibody production. Antigen-baited cytometry sorting enriched spike-specific memory B cells by ∼30-fold. Among 592 isolated mAbs, 53% bound the Wuhan spike, targeting the receptor-binding domain (28%), N-terminal domain (15%), or other regions (57%). Cross-reactivity analysis revealed that 40% of anti-spike mAbs recognized all tested variants of concern. VH/VL sequencing uncovered convergent rearrangements, including public V3-30 and V3-53/V3-66 clones, consistent with global findings. Two public receptor-binding domain-specific antibodies demonstrated broad neutralization when produced recombinantly. Together, these results validate the SCC system as a streamlined approach for unbiased repertoire analysis and functional mAb isolation. More broadly, the platform provides a practical framework for linking B cell clonal composition with antigen specificity and serum antibody responses. By reducing costs and simplifying workflows, it expands opportunities for antibody discovery and immunoepidemiological studies, fostering wider global participation in therapeutic antibody research.
Source: The Journal of Immunology
Authors: Luciana Conde, Debora L Oliveira, Gabriela Maciel, Fernando Castro, Aline de Oliveira Albuquerque, Danielle A Rodrigues, Yare Mëllo, Gustavo Meira de Assis, Bárbara Gabrielle, Suyane S Ferreira, Marcela S Cunha, Carlena Navas, Manuela C Emiliano, Marcele N Rocha, Barbara Soares, Lucas Tostes, Philippe Caloba, Bruno Maia, Francisco M Bastos de Oliveira, Amilcar Tanuri, Orlando C Ferreira, Terezinha M Castineiras, João Hermínio Martins da Silva, Juliana Echevarria, Marcelo T. Bozza, Leda R Castilho, Luciana J da Costa, Liza F Felicori, Alberto Nobrega, Gabriel D Victora, Carolina Lucas, Adriana Bonomo, André M Vale
Language: English
Subfield: Infectious Diseases
Field: Medicine
Domain: Health Sciences
Open Access status: green
Awards: ADEQUAÇÃO DO ESPAÇO BIOMAKER IDEAREAL COMO HUB DE ESPAÇOS MAKERS DA UFMG E PARA PROTOTIPAGEM E ESCALONAMENTO DE PRODUTOS, DESENVOLVIMENTO DE ESTRATÉGIAS COMPUTACIONAIS E EXPERIMENTAIS PARA ANÁLISE E PRODUÇÃO DE ANTICORPOS DE CAVALOS, INTEGRAÇÃO DE TECNOLOGIAS PARA DESCOBERTA E ANÁLISE FUNCIONAL DE ANTICORPOS VACINAIS CONTRA FEBRE AMARELA, G3077864592, DESENVOLVIMENTO DE UM PROCESSO PARA GERAÇÃO DE ANTICORPOS ANTI-SARS-COV-2 DE CAVALO
OpenAlex ID: W7117239908