AutoBlot: deterministic single-cell western blotting reveals proteomic diversity in rare cell populations

Single-cell western blotting (scWB) provides quantitative, molecular-mass-resolved protein measurements but relies on Poisson-limited gravity settling, requiring ~10 starting cells with <37% microwell occupancy. AutoBlot replaces stochastic loading with piezoelectric dispensing, achieving ~98% occupancy and deterministic cell-bead co-isolation from as few as 10,000 starting cells. Profiling patient-derived breast organoids (PDOs), we detect BRCA1-associated proteomic shifts, menopausal-status-dependent ER expression differences, and systematic differences between surface and intracellular lineage classification of mammary epithelial cells in organoid culture.

Deroy, C., Islam, M. N., Goldhammer, N., Jiang, L., Rosenbluth, J. M., Herr, A. E.

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