Optimization of the immunorecognition layer towards Brucella sp. on gold surface for SPR platform.

Autor: Pasquardini L; Indivenire srl, Via Sommarive 18, 38123 Trento, Italy. Electronic address: l.pasquardini@indiveni.re., Vanzetti L; Fondazione Bruno Kessler (FBK), Micro Nano Facility (MNF), Via Sommarive 18, 38123 Trento, Italy., Canteri R; Fondazione Bruno Kessler (FBK), Micro Nano Facility (MNF), Via Sommarive 18, 38123 Trento, Italy., Cennamo N; Department of Engineering, University of Campania 'L. Vanvitelli', Via Roma 29, 81031 Aversa, Italy., Arcadio F; Department of Engineering, University of Campania 'L. Vanvitelli', Via Roma 29, 81031 Aversa, Italy., Perri C; Moresense srl, Filarete Foundation, Viale Ortles 22/4, 20139 Milano, Italy., D'Agostino G; Moresense srl, Filarete Foundation, Viale Ortles 22/4, 20139 Milano, Italy., Pitruzzella R; Department of Engineering, University of Campania 'L. Vanvitelli', Via Roma 29, 81031 Aversa, Italy., Rovida R; Department of Engineering, University of Campania 'L. Vanvitelli', Via Roma 29, 81031 Aversa, Italy., Chiodi A; Moresense srl, Filarete Foundation, Viale Ortles 22/4, 20139 Milano, Italy., Zeni L; Department of Engineering, University of Campania 'L. Vanvitelli', Via Roma 29, 81031 Aversa, Italy. Electronic address: luigi.zeni@unicampania.it.
Jazyk: angličtina
Zdroj: Colloids and surfaces. B, Biointerfaces [Colloids Surf B Biointerfaces] 2023 Nov; Vol. 231, pp. 113577. Date of Electronic Publication: 2023 Oct 02.
DOI: 10.1016/j.colsurfb.2023.113577
Abstrakt: A successful immunosensor is characterized by a proper antibody immobilization and orientation in order to enhance the antigen recognition. In this work, a thorough characterization of the antibody functionalized gold surface is performed to set up the best conditions to implement in an optical platform for the detection of Brucella sp. Two different strategies are evaluated, based on a random immobilization and on an oriented one: a direct antibody immobilization on carboxylic mixed polyethylene (PEG) self-assembled monolayer (SAM) or only carboxylic PEG SAM interface is compared to an oriented immobilization on a layer of protein G on the same PEG SAM interfaces. X-ray Photoelectron Spectroscopy (XPS), Time of Flight Secondary Ion Mass Spectrometry (ToF-SIMS) and contact angle (CA) are used to chemically characterize the gold functionalized surface and ToF-SIMS is also used to confirm the right antibody orientation. Optical characterization is applied to monitor the functionalization steps and fluorescence measurements are used to set up the proper experimental conditions and also to detect Brucella bacteria on the surface. Best results are obtained with a 10 ng/μl incubation solution of antibody immobilized, in an oriented way, on a mixed PEG SAM interface.
Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
(Copyright © 2023 The Authors. Published by Elsevier B.V. All rights reserved.)
Databáze: MEDLINE
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