A more modest ~1.7-fold difference was obtained for anti-meningococcal vaccine hmAb cloning ~88% of hmAbs cloned via mICA versus ~53% cloned via the standard method were specific for a meningococcal surface protein. Following application of the modified whole-cell ICA (mICA), ~61% (19/31) of anti-pneumococcal polysaccharide hmAbs were cloned compared to 14% (8/59) obtained using standard (non-mICA) methods – representing a ~4.4-fold increase in hmAb cloning precision. Suspensions containing heterologous pneumococcal and meningococcal strains were then used to enrich for polysaccharide- and protein antigen-specific plasmablasts, respectively, during single cell sorting. Sequestration of IgG secreted by individual vaccine antigen-specific plasmablasts was achieved by the formation of an anti-CD45-streptavidin and biotin anti-IgG scaffold. Here, we report a novel modification of this single-antigen ICA using formalin-treated, fluorescently stained whole cell suspensions of the human bacterial invasive pathogens, Streptococcus pneumoniae and Neisseria meningitidis. Previously, a novel immunoglobulin-capture assay (ICA) was developed, using single protein vaccine antigens, to enhance the pathogen-specific hmAb cloning output. Precision of the hmAb cloning process relies on efficient isolation of hmAb-producing plasmablasts of interest. 3Section of Virology, Department of Infectious Disease, Imperial College London, London, United KingdomĮxpression cloning of fully human monoclonal antibodies (hmAbs) is seeing powerful utility in the field of vaccinology, especially for elucidating vaccine-induced B-cell responses and novel vaccine candidate antigen discovery. 2Flow Cytometry Core Facility, National Heart and Lung Institute, Imperial College London, London, United Kingdom.1Section of Paediatric Infectious Disease, Department of Infectious Disease, Imperial College London, London, United Kingdom.Gladstone 1†, Yanping Guo 2, Radhika Patel 2, Christopher L.
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