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

Abstract 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...

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Bibliographic Details
Published in:Nanomedicine 2015-02, Vol.11 (2), p.293-300
Main Authors: Ganau, Mario, MD, PhD, Bosco, Alessandro, PhD, Palma, Anita, PhD, Corvaglia, Stefania, Parisse, Pietro, PhD, Fruk, Ljiljana, PhD, Beltrami, Antonio Paolo, MD, Cesselli, Daniela, MD, Casalis, Loredana, PhD, Scoles, Giacinto, PhD
Format: Article
Language:English
Subjects:
Antibodies - chemistry
Antibodies - immunology
Antigens - chemistry
Antigens - immunology
Astrocytes - immunology
Astrocytes - pathology
Atomic force microscopy
Biomarkers - chemistry
Cancer biomarkers
DNA - chemistry
DNA-directed immobilization
Glial Fibrillary Acidic Protein - immunology
Glial Fibrillary Acidic Protein - isolation & purification
Glioma - diagnosis
Glioma - immunology
Humans
Immunoassay
Internal Medicine
Microscopy, Atomic Force
Nanodiagnostics
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Summary:Abstract 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.
ISSN:1549-9634
1549-9642
DOI:10.1016/j.nano.2014.04.006