Angela Maria Cusano
Istituto Italiano di Tecnologia
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Publication
Featured researches published by Angela Maria Cusano.
Journal of the American Chemical Society | 2015
Filippo Causa; Anna Aliberti; Angela Maria Cusano; Edmondo Battista; Paolo A. Netti
We present novel microgels as a particle-based suspension array for direct and absolute microRNA (miRNA) detection. The microgels feature a flexible molecular architecture, antifouling properties, and enhanced sensitivity with a large dynamic range of detection. Specifically, they possess a core-shell molecular architecture with two different fluorescent dyes for multiplex spectral analyses and are endowed with a fluorescent probe for miRNA detection. Encoding and detection fluorescence signals are distinguishable by nonoverlapping emission spectra. Tunable fluorescence probe conjugation and emission confinement on single microgels allow for ultrasensitive miRNA detection. Indeed, the suspension array has high selectivity and sensitivity with absolute quantification, a detection limit of 10(-15) M, a dynamic range from 10(-9) to 10(-15) M, and higher accuracy than qRT-PCR. The antifouling properties of the microgels also permit the direct measurement of miRNAs in serum, without sample pretreatment or target amplification. A multiplexed assay has been tested for a set of miRNAs chosen as cancer biomarkers.
Journal of the Royal Society Interface | 2014
Angela Maria Cusano; Filippo Causa; Raffaella Della Moglie; Nunzia Falco; Pasqualina Liana Scognamiglio; Anna Aliberti; Raffaele Vecchione; Edmondo Battista; Daniela Marasco; Marika Savarese; Umberto Raucci; Nadia Rega; Paolo A. Netti
In this paper, we report on a general approach for the detection of a specific tumoural biomarker directly in serum. Such detection is made possible using a protein-binding peptide selected through an improved phage display technique and then conjugated to engineered microparticles (MPs). Protein biomarkers represent an unlimited source of information for non-invasive diagnostic and prognostic tests; MP-based assays are becoming largely used in manipulation of soluble biomarkers, but their direct use in serum is hampered by the complex biomolecular environment. Our technique overcomes the current limitations as it produces a selective MP—engineered with an antifouling layer—that ‘captures’ the relevant protein staying impervious to the background. Our system succeeds in fishing-out the human tumour necrosis factor alpha directly in serum with a high selectivity degree. Our method could have great impact in soluble protein manipulation and detection for a wide variety of diagnostic applications.
International Journal of Biological Macromolecules | 2014
Yingli Liu; Angela Maria Cusano; Erin C. Wallace; Yasmina Mekmouche; Sana Ullah; Viviane Robert; Thierry Tron
Extremities of proteins are potent sites for functionalization. Carboxy terminus variants of the Trametes sp. strain C30 LAC3 laccase were generated and produced in Saccharomyces cerevisiae. A variant deleted of the last 13 residues (CΔ) and its 6 His tagged counterpart (CΔ6H) were found active enzymes. The production of CΔ6H resulted in the synthesis of a unusually high proportion of highly glycosylated forms of the enzyme therefore allowing the additional purification of a hyper-glycosylated form of CΔ6H noted CΔ6Hh. Properties of CΔ, CΔ6H and CΔ6Hh were compared. Globally, LAC3 catalytic efficiency was moderately affected by terminal modifications except in CΔ for which the kcat/KM ratio decreased 4 fold (with syringaldazine as substrate) and 10 fold (with ABTS as substrate) respectively. The catalytic parameters kcat and KM of CΔ6H and CΔ6Hh were found to be strictly comparable revealing that over glycosylation does not affect the enzyme catalytic efficiency. To the contrary, in vitro deglycosylation of laccase drastically reduced its activity. So, despite a complex glycosylated pattern observed for some of the variant enzymes, terminal sequences of laccases appear to be appropriate sites for the functionalization/immobilization of laccase.
Biomicrofluidics | 2016
David Dannhauser; Filippo Causa; Edmondo Battista; Angela Maria Cusano; Domenico Rossi; Paolo A. Netti
We present an in-flow ultrasensitive fluorescence detection of microRNAs (miRNAs) using spectrally encoded microgels. We researched and employed a viscoelastic fluid to achieve an optimal alignment of microgels in a straight measurement channel and applied a simple and inexpensive microfluidic layout, allowing continuous fluorescence signal acquisitions with several emission wavelengths. In particular, we chose microgels endowed with fluorescent emitting molecules designed for multiplex spectral analysis of specific miRNA types. We analysed in a quasi-real-time manner circa 80 microgel particles a minute at sample volumes down to a few microliters, achieving a miRNA detection limit of 202 fM in microfluidic flow conditions. Such performance opens up new routes for biosensing applications of particles within microfluidic devices.
Optical Methods for Inspection, Characterization, and Imaging of Biomaterials II | 2015
Filippo Causa; Anna Aliberti; Angela Maria Cusano; Edmondo Battista; Paolo A. Netti
Blood borne oligonucleotides fragments contain useful clinical information whose detection and monitoring represent the new frontier in liquid biopsy as they can transform the current diagnosis procedure. For instance, recent studies have identified a new class of circulating biomarkers such as s miRNAs, and demonstrated that changes in their concentration are closely associated with the development of cancer and other pathologies. However, direct detection of miRNAs in body fluids is particularly challenging and demands high sensitivity -concentration range between atto to femtomolarspecificity, and multiplexing Here we report on engineered multifunctional microgels and innovative probe design for a direct and multiplex detection of relevant clinical miRNAs in fluorescence by single particle assay. Polyethyleneglycol-based microgels have a coreshell architecture with two spectrally encoded fluorescent dyes for multiplex analyses and are endowed with fluorescent probes for miRNA detection. Encoding and detection fluorescence signals are distinguishable by not overlapping emission spectra. Tuneable fluorescence probe conjugation and corresponding emission confinement on single microgel allows for enhanced target detection. Such suspension array has indeed high selectivity and sensitivity with a detection limit of 10-15 M and a dynamic range from 10-9 to 10-15 M. We believe that sensitivity in the fM concentration range, signal background minimization, multiplexed capability and direct measurement of such microgels will translate into diagnostic benefits opening up new roots toward liquid biopsy in the context of point-of-care testing through an easy and fast detection of sensitive diagnostic biomarkers directly in serum.
Journal of Materials Science: Materials in Medicine | 2013
Nicola Gargiulo; Angela Maria Cusano; Filippo Causa; Domenico Caputo; Paolo A. Netti
Lab on a Chip | 2015
Francesco Del Giudice; Hojjat Madadi; Massimiliano M. Villone; Gaetano D'Avino; Angela Maria Cusano; Raffaele Vecchione; Maurizio Ventre; Pier Luca Maffettone; Paolo A. Netti
Analyst | 2016
Anna Aliberti; Angela Maria Cusano; Edmondo Battista; Filippo Causa; Paolo A. Netti
Proceedings of the Chemical Society, London | 1910
Anna Aliberti; Angela Maria Cusano; Edmondo Battista; Filippo Causa; Paolo A. Netti
Polymer International | 2016
Edmondo Battista; Alessia Mazzarotta; Filippo Causa; Angela Maria Cusano; Paolo A. Netti