Chemistries for Patterning Robust DNA MicroBarcodes Enable Multiplex Assays of Cytoplasm Proteins from Single Cancer Cells

摘要

The demand for parallel, multiplex analysis of protein biomarkers from ever smaller biospecimens is an increasing trend for both fundamental biology and clinical diagnostics.~([1–3]) The most highly multiplex protein assays rely on spatially encoded antibody microarrays,~([4–6]) and small biospecimens samples are now routinely manipulated using microfluidics approaches. The integration of antibody microarray techniques with microfluidics chips has only been explored relatively recently. One challenge arises from the relative instability of antibodies to microfluidics fabrication conditions. In recent years, several groups have devised methods to transform standard DNA microarrays in situ into protein microarrays and cell-capture platforms.~([7–13]) These approaches capitalize on the chemical robustness of DNA oligomers, and the reliable assembly of DNA-labeled structures via complementary hybridization. Recently, Fan et al. utilized a microfluidics-based flow patterning technique to generate DNA barcode-type arrays at 10x higher density than standard, spotted microarrays.~([14]) The DNA barcodes were converted into antibody arrays using the DNA-encoded antibody library (DEAL) technique, and then applied towards the measurement of a highly multiplex panel of proteins from a pinprick of whole blood.