SYSTEMS AND METHODS FOR MASSIVELY PARALLEL COMBINATORIAL ANALYSIS OF SINGLE CELLS

The present invention relates to an in vitro high-throughput cellular method for distinguishing reactive B cells from non-reactive B cells upon exposure to an antigen of interest comprising the steps of (a) extracting B cells from peripheral blood of a patient, (b) ex vivo incubation of the B cells with the antigen of interest inside aqueous-in-oil microdroplets or in nanoliter wells in a microfluidic device, (c) subjecting the B cells to emulsion overlap extension RT-PCR to generate a library of polynucleic acids that link heavy immunoglobulin sequences to transcripts being indicative of B cell activation, (d) identifying antibodies expressed by B cells that respond to the antigen of interest based on the activation biomarkers.