The United States Food and Drug Administration (FDA) approved carbohydrate antigen (CA)19-9 is not a reliable biomarker for the early detection of pancreatic ductal adenocarcinoma (PDAC).  Recent studies showed that circulating biomarkers such as cfDNA, microRNAs, and proteins in human plasma and/or serum can serve as more effective diagnostic markers for the early detection of pancreatic cancer. However, modern bioanalytical techniques such as quantitative reverse transcription polymerase chain reaction (qRT-PCR), Next Generation sequencing (NGS), microarray, lateral flow assay (LFA), and enzyme-linked immunosorbent assay (ELISA) are unable to assay different classes of biomolecules using a single instrument. For example, qRT-PCR and sequencing are used to assay only nucleic acids, whereas ELISA and LFA can detect only proteins. These techniques are also riddled with high occurrences of false positive responses, extremely labor intensive, need amplification and/or fluorescent labeling, and require extensive prior sample processing. Most importantly, assaying different classes of biomarkers and multiple markers of each type will improve the assay's specificity while mitigating false responses. Therefore, to overcome these challenges and improve the accuracy of the assay, we constructed a solid-state label-free nanosensor via covalent attachment of the photochromic molecule spiropyran to plasmonic gold triangular nanoprisms (Au TNPs). Exposure to UV light results in the formation of zwitterionic merocyanine surface that can bind either single stranded DNA or antibody receptors for microRNA or protein assays, respectively. Thus, our nanosensor can use an identical structural motif for highly specific detection of both microRNAs and proteins with sub-attomolar sensitivity.  The clinical utility of our nanosensor was demonstrated by detecting two microRNAs (microRNA-10b/155) and two proteins (CCL-20/CASP-8) in a cohort of 35 patients for early detection of PDAC. We also cross-validated the performance of our plasmonic nanosensor with droplet digital PCR and electrochemiluminescence method (mesoscale) for microRNA and protein assays, respectively.  We believe this highly innovative and customizable technology can be easily tuned to detect any current or future identified microRNA and/or protein biomarkers for the early detection of cancers and other diseases.

Adaptable plasmonic nanosensors comprised of photoswitchable molecular machines for multiparametric plasma biomarker detection facilitating diagnosis of pancreatic cancer

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