Abstract text: Every week, nearly 900 million washing machines operate globally - cleaning clothes predominantly made of cotton fibre. Collectively, these machines use ~19 billion m3 of water and emit the equivalent of ~62 million tonnes of CO2 each year. Most of the energy used by washing machines is consumed by water heating, so even modest reductions on washing temperatures have significant sustainability and cost benefits. Effective lower temperature washing is dependent on enzymes capable of breaking down soil components and/or disrupting their association with cotton fibre cell wall polymers. Further progress in developing enzyme-catalysed laundry formulations depends on deciphering the very complex molecular interactions operating at the cotton/soil interface – the ‘laundryome’. Our approach combines three technology platforms: High-throughput microarrays; molecular probes (antibodies and CBMs); and advanced microscopy. Here we show how these tools enable us to address key knowledge gaps that limit further development of enzyme-based laundry cleaning: what molecules are present in diverse soil types? How do they interact with each other, and with 3D cotton fibre cell wall architectures? What are the ‘keystone’ soil molecules that have disproportionally dominant roles in rendering soil complexes recalcitrant to removal? What enzyme classes show the most promise for deconstructing those complexes?