A DNA-based nano-immunoassay for the label-free detection of glial fibrillary acidic protein in multicell lysates

作者：Ganau Mario; Bosco Alessandro; Palma Anita; Corvaglia Stefania; Parisse Pietro; Fruk Ljiljana; Beltrami Antonio Paolo; Cesselli Daniela; Casalis Loredana^*; Scoles Giacinto

来源：Nanomedicine: Nanotechnology, Biology and Medicine , 2015, 11(2): 293-300.

DOI：10.1016/j.nano.2014.04.006

• Nanodiagnostics
• Cancer biomarkers
• Atomic force microscopy
• DNA-directed immobilization

We have developed a quantitative approach to eventually enable precise and multiplexing protein analysis of very small systems, down to a single or a few cells. Through DNA-directed immobilization of DNA-protein conjugates we immobilized antibodies specific for a certain protein of interest, on a complementary DNA nanoarray fabricated by means of nanografting, a nanolithography technique based on atomic force microscopy (AFM). The proof of concept was realized for glial fibrillary acidic protein (GFAP), a biomarker crucial in cell's differentiation of astrocytes, and functional to grade classification of gliomas, the most common of primary malignant brain tumors. The efficiency of the nano-immuno sensing was tested by obtaining the immobilization of purified recombinant GFAP protein at different concentration in a standard solution then in a cellular lysate. A comparison of sensitivity between our technique and conventional ELISA assays is provided at the end of the paper.
From the Clinical Editor: This team developed a quantitative approach to enable precise and multiplexing protein analysis of very small systems, down to a single or a few cells, demonstrating the utility of this DNA-based nano-immunoassay in the detection of GFAP.

• 出版日期2015-2