Article

Platelet-derived growth factor oncoprotein detection using three-dimensional carbon microarrays

Department of Mechanical and Materials Engineering, Florida International University, United States.
Biosensors & Bioelectronics (Impact Factor: 6.41). 07/2012; 39(1):118-23. DOI: 10.1016/j.bios.2012.06.055
Source: PubMed

ABSTRACT

The potential of aptamers as ligand binding molecule has opened new avenues in the development of biosensors for cancer oncoproteins. In this paper, a label-free detection strategy using signaling aptamer/protein binding complex for platelet-derived growth factor (PDGF-BB) oncoprotein detection is reported. The detection mechanism is based on the release of fluorophore (TOTO intercalating dye) from the target binding aptamer's stem structure when it captures PDGF. Amino-terminated three-dimensional carbon microarrays fabricated by pyrolyzing patterned photoresist were used as a detection platform. The sensor showed near linear relationship between the relative fluorescence difference and protein concentration even in the sub-nanomolar range with an excellent detection limit of 5pmol. This detection strategy is promising in a wide range of applications in the detection of cancer biomarkers and other proteins.

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    • "Therefore, the pyrolyzed carbon surface needs to be efficiently functionalized with different chemical groups including carboxylic group, amine group, sulfhydryl group, and hydroxyl group etc. in order to connect with biological materials such as recombinant antibodies, engineered proteins, and aptamers for the detection of wide variety of physiological substances. We have focused on developing various 3D micro/nanostructures using C-MEMS technique for the last decade [7] [8] [9] [10] [11] [12] [13] [14] [15] [16] [17] [18] [19]. In this study, different functionalization methods on C-MEMS based carbon microstructure have been investigated to form a highly selective, sensitive and reproducible miniaturized bio-sensing platform. "
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    ABSTRACT: Developing highly sensitive, selective, and reproducible miniaturized bio-sensing platforms require reliable biointerface which should be compatible with microfabrication techniques. In this study, we have fabricated pyrolyzed carbon arrays with high surface area as a bio-sensing electrode, and developed the surface functionalization methods to increase biomolecules immobilization efficiency and further understand electrochemical phenomena at biointerfaces. The carbon microelectrode arrays with high aspect ratio have been fabricated by carbon microelectromechanical systems (C-MEMS) and nanomaterials such as graphene have been integrated to further increase surface area. To achieve the efficient covalent immobilization of biomolecules, various oxidation and reduction functionalization methods have been investigated. The oxidation treatment in this study includes vacuum ultraviolet, electrochemical activation, UV/Ozone and oxygen RIE. The reduction treatment includes direct amination and diazonium grafting. The developed bio-sensing platform was then applied for several applications, such as: DNA sensor; H2O2 sensor; aptamer sensor and HIV sensor.
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    ABSTRACT: A novel method for the detection of PDGF-BB has been developed using double-strand DNA-copper nanoparticles (dsDNA-CuNPs) as fluorescent markers. This assay relies on the premise that the aptamer-based probe undergoes a conformational change upon binding with target protein, and subsequently triggers polymerization reaction to generate dsDNA. Then, the resultant dsDNA can be used as a template for the formation of CuNPs with high fluorescence. Under the optimized conditions, the proposed assay allowed sensitive and selective detection of PDGF-BB with a detection limit of 4 nmol/L. This possibly makes it an attractive platform for the detection of a variety of biomolecules whose aptamers undergo similar conformational change.
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    ABSTRACT: There is urgent need to develop highly selective, sensitive and reproducible miniaturized bio-sensing platform based on reliable interface that is compatible with microfabrication processing. Our research objective is to advance fundamental research by fabricating pyrolyzed carbon arrays with high surface area as a bio-sensing electrode, developing the functionalization methods to increasing biomolecules immobilization efficiency and further understanding electrochemical phenomena occurring at bio/carbon interfaces. The carbon microelectrode arrays with high aspect ratio and porous surface have been fabricated by carbon microelectromechanical systems (C-MEMS) while the nanomaterials such as graphene have been integrated to further increase surface area. To achieve the efficient covalent immobilization of biomolecules, various oxidation and reduction functionalization methods have been investigated. The oxidation treatment we used in this study includes vacuum ultraviolet, electrochemical activation, UV/Ozone and oxygen RIE. The reduction treatment includes direct amination and diazonium grafting. The functionalized surface has been characterized using XPS, CV and FTIR to confirm and calculate the surface coverage of different functional groups. The developed bio-sensing platform was then applied for several applications, such as: DNA sensor; Glucose sensor; H2O2 sensor; Aptamer sensor and HIV sensor. The performance and sensitivity of each biosensor will be discussed in the talk.
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