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Generating monoclonal antibodies from single B cells is a valuable tool for characterizing the specificity and functional properties of humoral responses. We and others developed protocols that have facilitated major advances in our understanding of B cell development, tolerance, and effector responses to HIV and influenza. Here, we demonstrate various refinements and dramatically reduce the time required to produce recombinant antibodies. Further, we present new methods for cloning and isolating antibodies from cells with lower immunoglobulin mRNA levels that may be resistant to traditional techniques. Together, these refinements significantly increase single-cell antibody expression efficiency and are easily integrated into established and novel pipelines. [ABSTRACT FROM AUTHOR]

The article analyzes the role of immunoglobulin A (IgA), the most abundant mammalian antibody isotype, in containing the resident commensal microbiota in the intestines. IgA is particularly prevalent at barrier surfaces and offers protection against enteric pathogens. Cloning of monoclonal antibodies (mAbs) from single murine IgA plasma cells is done to study the specificity of homeostatic IgA.