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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 \u223c30-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\u2013specific 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.<\/jats:p>","DOI":"10.1093\/jimmun\/vkaf305","type":"journal-article","created":{"date-parts":[[2025,12,25]],"date-time":"2025-12-25T05:47:36Z","timestamp":1766641656000},"source":"Crossref","is-referenced-by-count":2,"title":["Highly efficient and low-cost single-cell culture platform for unbiased analysis of human memory B cell repertoire and antibody discovery"],"prefix":"10.1093","volume":"215","author":[{"given":"Luciana","family":"Conde","sequence":"first","affiliation":[{"name":"Laborat\u00f3rio de Biologia de Linf\u00f3citos, Instituto de Biof\u00edsica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro , Rio de Janeiro, RJ,","place":["Brazil"]},{"name":"Department of Immunobiology, Yale University School of Medicine , New Haven, CT,","place":["USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Debora L","family":"Oliveira","sequence":"additional","affiliation":[{"name":"Laborat\u00f3rio de Biologia de Linf\u00f3citos, Instituto de Biof\u00edsica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro , Rio de Janeiro, RJ,","place":["Brazil"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gabriela","family":"Maciel","sequence":"additional","affiliation":[{"name":"Laborat\u00f3rio de Biologia de Linf\u00f3citos, Instituto de Biof\u00edsica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro , Rio de Janeiro, RJ,","place":["Brazil"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fernando","family":"Castro","sequence":"additional","affiliation":[{"name":"Laborat\u00f3rio de Biologia de Linf\u00f3citos, Instituto de Biof\u00edsica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro , Rio de Janeiro, RJ,","place":["Brazil"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Aline","family":"de Oliveira Albuquerque","sequence":"additional","affiliation":[{"name":"Funda\u00e7\u00e3o Oswaldo Cruz, Fiocruz Ceara, Pasteur-Fiocruz Center on Immunology and Immunotherapy , Eus\u00e9bio, CE,","place":["Brazil"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Danielle A","family":"Rodrigues","sequence":"additional","affiliation":[{"name":"Laborat\u00f3rio de Biologia de Linf\u00f3citos, Instituto de Biof\u00edsica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro , Rio de Janeiro, RJ,","place":["Brazil"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yare","family":"M\u00ebllo","sequence":"additional","affiliation":[{"name":"Laborat\u00f3rio de Biologia de Linf\u00f3citos, Instituto de Biof\u00edsica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro , Rio de Janeiro, 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Bozza","sequence":"additional","affiliation":[{"name":"Instituto de Microbiologia Paulo de G\u00f3es, Universidade Federal do Rio de Janeiro , Rio de Janeiro, RJ,","place":["Brazil"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Leda R","family":"Castilho","sequence":"additional","affiliation":[{"name":"Laborat\u00f3rio de Engenharia de Cultivos Celulares, Programa de Engenharia Qu\u00edmica, COPPE, Universidade Federal do Rio de Janeiro , Rio de Janeiro, RJ,","place":["Brazil"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Luciana J","family":"da Costa","sequence":"additional","affiliation":[{"name":"Instituto de Microbiologia Paulo de G\u00f3es, Universidade Federal do Rio de Janeiro , Rio de Janeiro, RJ,","place":["Brazil"]},{"name":"N\u00facleo de Enfrentamento e Estudos de Doen\u00e7as Infecciosas Emergentes e Reemergentes, Universidade Federal do Rio de Janeiro , Rio de Janeiro, 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