![]() A degree of clonal overlap was also observed between bone marrow and spleen antibody repertoires, indicating common origins of certain clones across lymphoid compartments. We observed that spleen B cell populations generated higher affinity antibodies compared to bone marrow PCs and that antigen-specific splenic B cells had higher average levels of somatic hypermutation. We performed affinity-based sorting of antibody yeast display libraries and large-scale next-generation sequencing analyses to follow antibody lineage performance, with experimental validation of 76 monoclonal antibodies against the cytokine antigen that identified three antibodies with exquisite double-digit picomolar binding affinity. To circumvent prior technical limitations for analysis of plasma cells, we applied single-cell antibody heavy and light chain gene capture from the entire mature B cell repertoires followed by yeast display functional analysis using a cytokine as a model immunogen. Here, we studied the antibodies encoded by the entire mature B cell populations, including PCs, and compared the antibody repertoires of bone marrow and spleen compartments elicited by immunization in a human immunoglobulin transgenic mouse strain. Despite the importance of plasma cells (PCs) as the dominant source of circulating antibodies in serum, PCs are rarely utilized because they do not express surface BCRs and cannot be analyzed using antigen-based fluorescence-activated cell sorting. Molecular characterization of antibody immunity and human antibody discovery is mainly carried out using peripheral memory B cells, and occasionally plasmablasts, that express B cell receptors (BCRs) on their cell surface. 8Department of Chemical Engineering, The University of Kansas, Lawrence, KS, United States.7Sanofi Vaccines, Sanofi, Cambridge, MA, United States.6Large Molecule Research, Sanofi, Cambridge, MA, United States.5Large Molecule Research, Sanofi, Ghent, Belgium.4Large Molecule Research, Sanofi, Vitry sur Seine, France.3Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, MA, United States.2Ragon Institute of Massachusetts General Hospital (MGH), Massachusetts Institute of Technology (MIT), and Harvard, Cambridge, MA, United States. ![]() 1Department of Pharmaceutical Chemistry, The University of Kansas, Lawrence, KS, United States.Gutiérrez-González 2,3 Matheus Oliveira de Souza 1,2,3 Joshua M. Rao 6 David Aguilera-Rodríguez 5 Mathilde Rezé 4 Christine A. López Acevedo 1 Camille Cuziol 4 Evelyn De Tavernier 5 Ahmed S. ![]()
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