Di Natale, Concetta (2016) Miniaturized biosensors and microdevices based on peptide for food industry and healthcare. [Tesi di dottorato]

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Item Type: Tesi di dottorato
Lingua: English
Title: Miniaturized biosensors and microdevices based on peptide for food industry and healthcare
Creators:
CreatorsEmail
Di Natale, Concettaconny_dn88@hotmail.it
Date: 31 March 2016
Number of Pages: 136
Institution: Università degli Studi di Napoli Federico II
Department: Ingegneria Chimica, dei Materiali e della Produzione Industriale
Scuola di dottorato: Ingegneria industriale
Dottorato: Ingegneria dei materiali e delle strutture
Ciclo di dottorato: 28
Coordinatore del Corso di dottorato:
nomeemail
Mensitieri, Giuseppegiuseppe.mensitieri@unina.it
Tutor:
nomeemail
Causa, FilippoUNSPECIFIED
Netti, Paolo AntonioUNSPECIFIED
Date: 31 March 2016
Number of Pages: 136
Uncontrolled Keywords: biosensors;microdevices;peptides;microgels;food industry;healthcare
Settori scientifico-disciplinari del MIUR: Area 09 - Ingegneria industriale e dell'informazione > ING-IND/22 - Scienza e tecnologia dei materiali
Date Deposited: 13 Apr 2016 00:10
Last Modified: 04 May 2017 01:00
URI: http://www.fedoa.unina.it/id/eprint/11013

Abstract

Molecular recognition events are some of the most significant features of biological and chemical systems. The biomolecular recognition is the ability of a biomolecule to interact selectively with another molecule even in the presence of structurally similar antagonist molecules. Molecular recognition is fundamental for biosensing technique. A biosensor, according to IUPAC recommendations 1999, is an independently integrated receptor transducer device, which is capable of providing selective quantitative or semi-quantitative analytical information using a biological recognition element. A biosensor consists of three main elements, a bioreceptor, a transducer, and a signal processing system.The bioreceptor or biological recognition element is the most important and significant feature of a biosensor. The bioreceptor is the heart of the recognition system of a sensor towards the target analyte. Essentially it is crucial for a bioreceptor to be selective and sensitive towards the specific target in order to prevent the interference by other substance from sample matrix. Peptides have been used as components in biological analysis and fabrication of novel biosensors for a number of reasons, including mature synthesis protocols, highly selectivity for enzymes substrates and low production costs. Bio-conjugation strategies can provide an efficient way to convert interaction information between peptides and analytes into a measurable signal, which can be used for fabrication of novel peptide-based biosensors. Furthermore, peptides based biosensors are presented with a description of their properties, features and advantages. The use of peptides as capture agents and the possibility to exploit their reversibility in biological applications, despite proteins and antibodies, is still very limited and has not been addressed yet. This aspect has been investigated in this first chapter. In Chapter 2 microfluidic synthesis of novel polymeric microparticles endowed with specific peptide due to its superior specificity for target binding in complex media, is reported. In more details, a peptide sequence is efficiently encapsulated into the polymeric network and protein binding occurred with high affinity (KD 0.1-0.4 µM). Fluidic dynamics simulation is performed to optimize the production conditions for monodisperse and stable functionalized microgels. The results demonstrate the easy and fast realization, in a single step, of functionalized monodisperse microgels using droplet-microfluidic technique, and how the inclusion of the peptide within polymeric network improves both the affinity and the specificity of protein capture. In Chapter 3 the development of a peptide-based biosensor for autofluorescence aflatoxin M1 detection in a sensitive, specific and unsophisticated manner, is reported. To this aim an integrated approach has been developed to select specific peptide motif to capture aflatoxin M1. The integrated approach provides a combination of computation modeling with combinatorial peptide synthesis to screen the sequence with the highest affinity. Peptides sequences selected by the proposed approach have been easily co-polymerized in PEGDA microparticles opening the route towards a direct detection of aflatoxins in small volume both in liquid and solid environments. Such approach can be applied also to other small molecules to develop materials able to sequestrate the target analytes allowing their direct detection directly in the materials. Finally in Chapter 4 a microdevice for endometriosis diagnosis is developed. In this work, three different peptides (CRP-1, VEGF-114 and ΦG6) are used as capture agents to detect serum levels of vascular endothelial growth factor (VEGF), tumor necrosis factor-alpha (TNF-a), and C-reactive protein (CRP): three serum markers of endometriosis in menstrual blood. The selected peptides have been covalently immobilized on a microfluidic PDMS device, previously derivatized with 10% of PAA (Poly(acrylic acid)) solution. The so built device has been used to capture and recognize endometriosis markers both in buffer and biologic fluids matrices such as human serum with a good specificity and sensitivity. So the aim of the studies in this work, is to set up miniaturized microdevices that can be used in healthcare and food industry, using peptides as capture agents in order to have sensitive but not invasive and not expansive biosensors.

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