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Paper-Based Immunosensors with Bio-Chemiluminescence Detection

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Paper-Based Immunosensors with Bio-Chemiluminescence Detection sensors Review Paper-Based Immunosensors with Bio-Chemiluminescence Detection Maria Maddalena Calabretta 1,2,† , Martina Zangheri 1,†, Donato Calabria 1 , Antonia Lopreside 1,2, Laura Montali 1,2, Elisa Marchegiani 1, Ilaria Trozzi 1, Massimo Guardigli 1,3, Mara Mirasoli 1,3,4,* and Elisa Michelini 1,2,4,5,* 1 Department of Chemistry “Giacomo Ciamician”, University of Bologna, 40126 Bologna, Italy; [email protected] (M.M.C.); [email protected] (M.Z.); [email protected] (D.C.); [email protected] (A.L.); [email protected] (L.M.); [email protected] (E.M.); [email protected] (I.T.); [email protected] (M.G.) 2 Center for Applied Biomedical Research (CRBA), University of Bologna, 40138 Bologna, Italy 3 Interdepartmental Centre for Renewable Sources, Environment, Sea and Energy (CIRI FRAME), Alma Mater Studiorum, University of Bologna, 48123 Ravenna, Italy 4 INBB, Istituto Nazionale di Biostrutture e Biosistemi, Via Medaglie d’Oro, 00136 Rome, Italy 5 Health Sciences and Technologies-Interdepartmental Center for Industrial Research (HST-ICIR), University of Bologna, 40126 Bologna, Italy * Correspondence: [email protected] (M.M.); [email protected] (E.M.) † These authors contributed equally. Abstract: Since the introduction of paper-based analytical devices as potential diagnostic platforms a few decades ago, huge efforts have been made in this field to develop systems suitable for meeting the requirements for the point-of-care (POC) approach. Considerable progress has been achieved in the adaptation of existing analysis methods to a paper-based format, especially considering the Citation: Calabretta, M.M.; Zangheri, chemiluminescent (CL)-immunoassays-based techniques. The implementation of biospecific assays M.; Calabria, D.; Lopreside, A.; with CL detection and paper-based technology represents an ideal solution for the development of Montali, L.; Marchegiani, E.; Trozzi, I.; Guardigli, M.; Mirasoli, M.; Michelini, portable analytical devices for on-site applications, since the peculiarities of these features create E. Paper-Based Immunosensors with a unique combination for fitting the POC purposes. Despite this, the scientific production is not Bio-Chemiluminescence Detection. paralleled by the diffusion of such devices into everyday life. This review aims to highlight the open Sensors 2021, 21, 4309. https:// issues that are responsible for this discrepancy and to find the aspects that require a focused and doi.org/10.3390/s21134309 targeted research to make these methods really applicable in routine analysis. Academic Editor: Huangxian Ju Keywords: paper-based; bio-chemiluminescence; immunosensor; biosensor; point-of-care Received: 23 May 2021 Accepted: 21 June 2021 Published: 24 June 2021 1. Introduction Immunoassays have been routinely used in laboratories equipped with bulky in- Publisher’s Note: MDPI stays neutral strumentation and skilled personnel for the quantification of target analytes for various with regard to jurisdictional claims in applications in healthcare, food safety and environmental monitoring. In recent years, published maps and institutional affil- iations. research has mainly focused on making the immunoassay technique suitable for portable analytical formats, to perform low-cost tests directly on site [1]. Indeed, thanks to their selectivity and specificity, immunoassays allow the detection of analytes within complex matrices without the need for long-processing sample pretreatments. Among portable for- mats suitable for the development of point-of-care (POC) immunoassays, the paper-based Copyright: © 2021 by the authors. approach has attracted strong interest thanks to its advantageous features, including low Licensee MDPI, Basel, Switzerland. cost, straightforward procedures, flexibility of the paper-based support, short turnaround This article is an open access article time and small consumption of samples and reagents. For these reasons, paper-based POC distributed under the terms and conditions of the Creative Commons immunoassays are expected to become a feasible option for monitoring human health- Attribution (CC BY) license (https:// care, food safety or water quality in resource-limited settings and, thanks to the recent creativecommons.org/licenses/by/ improvements in microfluidics and nanotechnologies, they have developed rapidly in 4.0/). recent years [1]. Among the different varieties of paper-based format biosensors, those Sensors 2021, 21, 4309. https://doi.org/10.3390/s21134309 https://www.mdpi.com/journal/sensors Sensors 2021, 21, x FOR PEER REVIEW 2 of 18 Sensors 2021, 21, 4309 2 of 18 biosensors, those combined most frequently with immunoassays are lateral-flow immu- noassayscombined (LFIA) mostand microfluidic frequently with paper immunoassays-based devices are (µPADs) lateral-flow [2]. immunoassays (LFIA) and LFIAmicrofluidic is an integrated paper-based platform devices in which (µPADs) pre [2-].stored reagents are entrapped on a ni- trocelluloseLFIA strip. is By an adding integrated the sample, platform these in reagents which pre-stored are solubilized reagents and, arewhile entrapped they are on a flowingnitrocellulose through the strip.membrane, By adding bioassays the take sample, place. these On the reagents surface are of solubilizedthe nitrocellulose and, while strip, theydifferent are flowingareas contain through specific the membrane,probes responsible bioassays for the take recognition place. On and the detection surface of the of thenitrocellulose target analyte. strip, It is differentpossible areasto exploit contain different specific bioassay probes responsibleformats, including for the recognitionsand- wich, andcompetitive detection or ofmultiplex the target formats analyte.. It is possible to exploit different bioassay formats, Asincluding an alternative, sandwich, μPADs competitive are characterized or multiplex by formats.the presence of hydrophilic and hy- µ drophobic Asmicrocha an alternative,nnels, whichPADs allow are to characterized develop different by the designs presence suitable of hydrophilic for the de- and hy- sired drophobicapplication. microchannels, The microfluidic which pattern allow is to generally develop different obtained designs by exploiting suitable chemical for the desired printingapplication. and/or cutting The microfluidic and it is possible pattern to is develop generally 2D obtained or 3D configuration by exploitings chemicalfor moving printing and/or cutting and it is possible to develop 2D or 3D configurations for moving fluids fluids through vertical and horizontal pathways [2,3]. through vertical and horizontal pathways [2,3]. The analytical performance of miniaturized analytical devices represents the bottle- The analytical performance of miniaturized analytical devices represents the bottle- neck when high sensitivity is required, such as for the detection of analytes present at neck when high sensitivity is required, such as for the detection of analytes present at trace trace levels in complex matrices [4,5]. For this reason, one of the most important aspects levels in complex matrices [4,5]. For this reason, one of the most important aspects in the in the development of these methods is the selection of the detection technique since it has development of these methods is the selection of the detection technique since it has to to combine high sensitivity with reduced sensing equipment, providing the possibility of combine high sensitivity with reduced sensing equipment, providing the possibility of working with portable and low-cost devices and ease of operation even by non-special- working with portable and low-cost devices and ease of operation even by non-specialized ized personnel [5]. Several attempts to avail popular detection methods. such as UV/Vis personnel [5]. Several attempts to avail popular detection methods. such as UV/Vis absorp- absorption spectroscopy [6–9], have been reported in literature but the most promising tion spectroscopy [6–9], have been reported in literature but the most promising techniques techniques are those based on electrochemical and luminescence detection. are those based on electrochemical and luminescence detection. In particular,In particular, chemical chemical luminescence luminescence is based is based on the on production the production of photons of photons triggered triggered by a chemicalby a chemical reaction, reaction, as for as chemiluminescence for chemiluminescence (CL), (CL),and, when and, whenthe reaction the reaction occurs occurs withinwithin living living organisms, organisms, the phe thenomenon phenomenon is called is called bioluminescence bioluminescence (BL). (BL).Since SinceBL and BL and CL reactionCL reactionss start in start the dark, in the photons dark, photons can be measured can be measured with high with efficiency high efficiency ensuring ensuringthe absencethe of absence nonspecific of nonspecific signals, signals,thus avoiding thus avoiding the background the background commonly commonly encountered encountered with photoluminescencewith photoluminescence measurem measurements.ents. For this For reason, this reason, CL-based CL-based analytical analytical methods methods can can achieveachieve,, in principle in principle,, high highdetectability detectability and andcan represent can represent an alternative an alternative or a or complemen- a complementary tary approachapproach in in the the field field of of optical optical biosensors biosensors [10]. [10]. TogetherTogether with withtheirtheir
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