METHOD OF SCREENING A PLURALITY OF SINGLE SECRETING CELLS FOR FUNCTIONAL ACTIVITY

摘要

This invention generally relates to methods, devices and kits for screening a plurality of single secreting cells for functional activity of the secreted molecules by measuring the amount of reporter gene mRNA produced in one or more reporter cells in response to the secreted molecules.

主权项

1. A method of measuring the functional activity of a secreted molecule secreted by a secreting cell consisting of:
placing a plurality of secreting cells into microwells such that a single secreting cell occupies a single microwell and is secreting molecules; placing one or more reporter cells of one or more types into microwells; allowing copies of the secreted molecules to interact with a receptor of the reporter cells; capturing mRNA from one or more of the reporter genes of the reporter cells and capturing mRNA from one or more of the housekeeping genes of the reporter cells with an oligonucleotide capture support containing mRNA capture oligos on each feature comprising a unique DNA tag; converting captured mRNA into cDNA incorporating the DNA tag; sequencing the tagged and optionally coded cDNA by NGS; examining the sequenced cDNA from the one or more reporter genes and comparing it to the sequenced cDNA from the one or more housekeeping genes of the reporter cells to determine the functional activity of a secreted molecule secreted by a secreting cell.