High-throughput identification of intermolecular interactions

A method of detecting an interaction between a macromolecule and an interaction partner, the method comprising:

• obtaining a composition comprising a plurality of macromolecules and a plurality of candidate interaction partners;
• performing two or more iterations of split-and-pool barcoding, each iteration comprising:
• (i) randomly distributing the composition into a plurality of partitions in fluid isolation from each other;
• (ii) in the plurality of partitions, barcoding the plurality of macromolecules and the plurality of candidate interaction partners with a combinatorial barcode unit,
• wherein within each partition, the plurality of macromolecules and the plurality of candidate interaction partners are barcoded with the same combinatorial barcode unit,
• wherein the plurality of macromolecules and the plurality of candidate interaction partners of different partitions receive different combinatorial barcode units from each other
• wherein a macromolecule and a candidate interaction partner that interact will assort together in a partition of the plurality of partitions;
• and
• (iii) pooling the barcoded plurality of macromolecules and the barcoded plurality of candidate interaction partners comprising the different combinatorial barcode units from the plurality of partitions into a single composition,
• wherein, after the two or more iterations of split-and-pool barcoding, each macromolecule of the plurality of macromolecules and each candidate interaction partner of the plurality of candidate interaction partners comprises a combinatorial barcode comprising two or more combinatorial barcode units, each combinatorial barcode unit corresponding to an iteration of split-and-pool-barcoding;
• and
• obtaining identifying information from the combinatorial barcodes of the plurality of macromolecules and the plurality of candidate interaction partners, wherein the identifying information permits detecting an interaction between a macromolecule of the plurality of macromolecules and a candidate interaction partner of the plurality of candidate interaction partners through the two or more iterations of split-and-pool barcoding,
• wherein a probability of the interaction between the macromolecule and the candidate interaction partner as being bona fide is proportional to the number of iterations of split-and-pool barcoding.