NAPPA based nanogravimetric biosensor: Preliminary characterization

Spera, Rosanna; Terencio, Tercio Bezerra Correia; Nicolini, Claudio

doi:10.1016/j.snb.2013.03.063

Cited by 12 publications

(32 citation statements)

References 25 publications

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“…In SNAP-NAPPA microarrays the proteins are synthesized with the addition of self-labeling SNAP tag, which is a 20 kDa mutant of DNA repair protein that reacts with benzylguanine derivatives [61,62] leading to irreversible covalent labeling of SNAP tag, which is used for protein immobilization [63]. Many detection approaches, such as fluorescence and enzymatic methods, but also label-free nanotechnology-based techniques such as atomic force microscopy (AFM), nanogravimetry [64], quartz crystal microbalance (QCM), anodic porous alumina (APA) or mass spectrometry (for description of detection techniques see section 'Detection techniques employed in nanotechnologies-based protein arrays') can be employed. To our best knowledge, to this date NAPPA is the only c ommercially used t echnology based on nanosystems [65].…”

Section: In Situ Protein Microarraysmentioning

confidence: 99%

Nanotechnologies in Protein Microarrays

Křížková

Heger

Zalewska

et al. 2015

Nanomedicine (Lond.)

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Protein microarray technology became an important research tool for study and detection of proteins, protein-protein interactions and a number of other applications. The utilization of nanoparticle-based materials and nanotechnology-based techniques for immobilization allows us not only to extend the surface for biomolecule immobilization resulting in enhanced substrate binding properties, decreased background signals and enhanced reporter systems for more sensitive assays. Generally in contemporarily developed microarray systems, multiple nanotechnology-based techniques are combined. In this review, applications of nanoparticles and nanotechnologies in creating protein microarrays, proteins immobilization and detection are summarized. We anticipate that advanced nanotechnologies can be exploited to expand promising fields of proteins identification, monitoring of protein-protein or drug-protein interactions, or proteins structures.

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Section: In Situ Protein Microarraysmentioning

confidence: 99%

Nanotechnologies in Protein Microarrays

Křížková

Heger

Zalewska

et al. 2015

Nanomedicine (Lond.)

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“…We also tested the possibility to analyze drug/small molecule-protein interactions in QC displaying multiple proteins, a task which is not possible with fluorescence based arrays (Spera et al, 2013a(Spera et al, ,2013b.…”

Section: Nucleic Acid Programmable Protein Arraysmentioning

confidence: 99%

“…The calibration curves equation (obtained with Ordinary Least Squares methods, OLS) are (Spera et al, 2013a(Spera et al, ,2013b: The calibration curves equation (obtained with Ordinary Least Squares methods, OLS) are (Spera et al, 2013a(Spera et al, ,2013b:…”

Section: Nucleic Acid Programmable Protein Arraysmentioning

confidence: 99%

Proteomics and Proteogenomics Approaches for Oral Diseases

Bragazzi

Pechkova

Nicolini

2014

Advances in Protein Chemistry and Structural Biology

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“…[3]; the labeled molecule is then detected via a fluorescence microscope, flow cytometer or some other fluorescence reading instrument [4]. On the contrary label-free analysis do not require the use of reporter elements (fluorescent, luminescent, radiometric, or colorimetric) to facilitate measurements, it can provide direct information on analyte binding to array molecules typically in the form of mass addition or depletion from the array surface [5][6][7].…”

Section: Introductionmentioning

confidence: 99%

Mass Spectrometry and Florescence Analysis of Snap-Nappa Arrays Expressed Using E. coli Cell_Free Expression System

Nicolini

Spera

Festa³

et al. 2013

J Nanomed Nanotechnol

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We present the analysis of an innovative kind of self-assembling protein microarray, the "Nucleic Acid Programmable Protein Array" (NAPPA), express with the SNAP tag in E.coli coupled self free expression system. The goal is to develop a standardize procedure to analyze the protein protein interaction occurred on NAPPA array combining label free Mass Spectrometry (MS) and fluorescence technology for protein microarray. We employ in the process "Protein synthesis Using Recombinant Elements" (PURE) system. For the first time an improved version of NAPPA, that allows for functional proteins to be synthesized in situ -with a SNAP tag -directly from printed cDNAs just in time for assay, has been expressed with a novel cell-free transcription/translation system reconstituted from the purified components necessary for Escherichia coli translation -the PURE system -and analyzed both in fluorescence and in a label free manner by four different mass spectrometers, namely three Matrix Assisted Laser Desorption Ionization Time-of-Flight (MALDI-TOF), a Voyager, a Bru ker Autoflex and a Bruker Ultraflex, and Liquid Chromatography-Electrospray Ionization Mass Spectrometry (LC-ESI-MS/MS). Due to the high complexity of the system, an ad hoc bioinformatic tool has been needed to develop for their successful analysis. The contemporary fluorescence analysis of NAPPA, expressed by means of PURE system, has been performed to confirm the improved characterization of this new NAPPA-SNAP system.

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NAPPA based nanogravimetric biosensor: Preliminary characterization

Cited by 12 publications

References 25 publications

Nanotechnologies in Protein Microarrays

Nanotechnologies in Protein Microarrays

Proteomics and Proteogenomics Approaches for Oral Diseases

Mass Spectrometry and Florescence Analysis of Snap-Nappa Arrays Expressed Using E. coli Cell_Free Expression System

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