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Prof. Department Chemistry
Dipartimento di Chimica, Università degli Studi di Torino

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Research Keywords & Expertise

0 Bioanalytical Chemistry
0 Immunoassays
0 Lateral Flow Assays
0 biosensing
0 enzyme immunoassay

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Immunoassays
enzyme immunoassay
biosensing

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Short Biography

Laura Anfossi is an associate professor at the Department of Chemistry, University of Turin, and she works in the Forensic, Analytical and Bioanalytical Laboratories. She has been working for several years on the development of immunological methods of analysis in several formats and for disparate applications (food safety, clinics, veterinary). Her research interest is focused on new strategies and new materials for point-of-need tests based on lateral flow assay technology.

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Journal article
Published: 10 August 2021 in Polymers
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An innovative approach to imprinted nanoparticles (nanoMIPs) is represented by solid-phase synthesis. Since the polymeric chains grow over time and rearrange themselves around the template, the binding properties of nanoMIPs could depend on the polymerization time. Here we present an explorative study about the effect of different polymerization times on the binding properties of ciprofloxacin-imprinted nanoMIPs. The binding properties towards ciprofloxacin were studied by measuring the binding affinity constants (Keq) and the kinetic rate constants (kd, ka). Furthermore, selectivity and nonspecific binding were valued by measuring the rebinding of levofloxacin onto ciprofloxacin-imprinted nanoMIPs and ciprofloxacin onto diclofenac-imprinted nanoMIPs, respectively. The results show that different polymerization times produce nanoMIPs with different binding properties: short polymerization times (15 min) produced nanoMIPs with high binding affinity but low selectivity (Keq > 107 mol L−1, α ≈ 1); medium polymerization times (30 min–2 h) produced nanoMIPs with high binding affinity and selectivity (Keq ≥ 106 mol L−1, α < 1); and long polymerization times (>2 h) produced nanoMIPs with low binding affinity, fast dissociation kinetics and low selectivity (Keq ≤ 106 mol L−1, kdis > 0.2 min−1, α ≈ 1). The results can be explained as the combined effect of rearrangement and progressive stiffening of the polymer chains around the template molecules.

ACS Style

Matteo Chiarello; Laura Anfossi; Simone Cavalera; Fabio Di Nardo; Fiora Artusio; Roberto Pisano; Claudio Baggiani. Effect of Polymerization Time on the Binding Properties of Ciprofloxacin-Imprinted nanoMIPs Prepared by Solid-Phase Synthesis. Polymers 2021, 13, 2656 .

AMA Style

Matteo Chiarello, Laura Anfossi, Simone Cavalera, Fabio Di Nardo, Fiora Artusio, Roberto Pisano, Claudio Baggiani. Effect of Polymerization Time on the Binding Properties of Ciprofloxacin-Imprinted nanoMIPs Prepared by Solid-Phase Synthesis. Polymers. 2021; 13 (16):2656.

Chicago/Turabian Style

Matteo Chiarello; Laura Anfossi; Simone Cavalera; Fabio Di Nardo; Fiora Artusio; Roberto Pisano; Claudio Baggiani. 2021. "Effect of Polymerization Time on the Binding Properties of Ciprofloxacin-Imprinted nanoMIPs Prepared by Solid-Phase Synthesis." Polymers 13, no. 16: 2656.

Review
Published: 30 July 2021 in Sensors
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The Lateral Flow Immunoassay (LFIA) is by far one of the most successful analytical platforms to perform the on-site detection of target substances. LFIA can be considered as a sort of lab-in-a-hand and, together with other point-of-need tests, has represented a paradigm shift from sample-to-lab to lab-to-sample aiming to improve decision making and turnaround time. The features of LFIAs made them a very attractive tool in clinical diagnostic where they can improve patient care by enabling more prompt diagnosis and treatment decisions. The rapidity, simplicity, relative cost-effectiveness, and the possibility to be used by nonskilled personnel contributed to the wide acceptance of LFIAs. As a consequence, from the detection of molecules, organisms, and (bio)markers for clinical purposes, the LFIA application has been rapidly extended to other fields, including food and feed safety, veterinary medicine, environmental control, and many others. This review aims to provide readers with a 10-years overview of applications, outlining the trends for the main application fields and the relative compounded annual growth rates. Moreover, future perspectives and challenges are discussed.

ACS Style

Fabio Di Nardo; Matteo Chiarello; Simone Cavalera; Claudio Baggiani; Laura Anfossi. Ten Years of Lateral Flow Immunoassay Technique Applications: Trends, Challenges and Future Perspectives. Sensors 2021, 21, 5185 .

AMA Style

Fabio Di Nardo, Matteo Chiarello, Simone Cavalera, Claudio Baggiani, Laura Anfossi. Ten Years of Lateral Flow Immunoassay Technique Applications: Trends, Challenges and Future Perspectives. Sensors. 2021; 21 (15):5185.

Chicago/Turabian Style

Fabio Di Nardo; Matteo Chiarello; Simone Cavalera; Claudio Baggiani; Laura Anfossi. 2021. "Ten Years of Lateral Flow Immunoassay Technique Applications: Trends, Challenges and Future Perspectives." Sensors 21, no. 15: 5185.

Review
Published: 12 May 2021 in Sensors
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Paper-based lateral-flow immunoassays (LFIAs) have achieved considerable commercial success and their impact in diagnostics is continuously growing. LFIA results are often obtained by visualizing by the naked eye color changes in given areas, providing a qualitative information about the presence/absence of the target analyte in the sample. However, this platform has the potential to provide ultrasensitive quantitative analysis for several applications. Indeed, LFIA is based on well-established immunological techniques, which have known in the last year great advances due to the combination of highly sensitive tracers, innovative signal amplification strategies and last-generation instrumental detectors. All these available progresses can be applied also to the LFIA platform by adapting them to a portable and miniaturized format. This possibility opens countless strategies for definitively turning the LFIA technique into an ultrasensitive quantitative method. Among the different proposals for achieving this goal, the use of enzyme-based immunoassay is very well known and widespread for routine analysis and it can represent a valid approach for improving LFIA performances. Several examples have been recently reported in literature exploiting enzymes properties and features for obtaining significative advances in this field. In this review, we aim to provide a critical overview of the recent progresses in highly sensitive LFIA detection technologies, involving the exploitation of enzyme-based amplification strategies. The features and applications of the technologies, along with future developments and challenges, are also discussed.

ACS Style

Donato Calabria; Maria Calabretta; Martina Zangheri; Elisa Marchegiani; Ilaria Trozzi; Massimo Guardigli; Elisa Michelini; Fabio Di Nardo; Laura Anfossi; Claudio Baggiani; Mara Mirasoli. Recent Advancements in Enzyme-Based Lateral Flow Immunoassays. Sensors 2021, 21, 3358 .

AMA Style

Donato Calabria, Maria Calabretta, Martina Zangheri, Elisa Marchegiani, Ilaria Trozzi, Massimo Guardigli, Elisa Michelini, Fabio Di Nardo, Laura Anfossi, Claudio Baggiani, Mara Mirasoli. Recent Advancements in Enzyme-Based Lateral Flow Immunoassays. Sensors. 2021; 21 (10):3358.

Chicago/Turabian Style

Donato Calabria; Maria Calabretta; Martina Zangheri; Elisa Marchegiani; Ilaria Trozzi; Massimo Guardigli; Elisa Michelini; Fabio Di Nardo; Laura Anfossi; Claudio Baggiani; Mara Mirasoli. 2021. "Recent Advancements in Enzyme-Based Lateral Flow Immunoassays." Sensors 21, no. 10: 3358.

Paper
Published: 18 November 2020 in Analytical Methods
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A selective molecularly imprinted polymer prepared with a squaramide-based functional monomer was used for the solid phase extraction of roxarsone from surface waters.

ACS Style

Simone Cavalera; Fabio Di Nardo; Giulia Spano; Laura Anfossi; Panagiotis Manesiotis; Claudio Baggiani. Stoichiometric molecular imprinting using polymerisable urea and squaramide receptors for the solid phase extraction of organo-arsenic compound roxarsone. Analytical Methods 2020, 12, 5729 -5736.

AMA Style

Simone Cavalera, Fabio Di Nardo, Giulia Spano, Laura Anfossi, Panagiotis Manesiotis, Claudio Baggiani. Stoichiometric molecular imprinting using polymerisable urea and squaramide receptors for the solid phase extraction of organo-arsenic compound roxarsone. Analytical Methods. 2020; 12 (47):5729-5736.

Chicago/Turabian Style

Simone Cavalera; Fabio Di Nardo; Giulia Spano; Laura Anfossi; Panagiotis Manesiotis; Claudio Baggiani. 2020. "Stoichiometric molecular imprinting using polymerisable urea and squaramide receptors for the solid phase extraction of organo-arsenic compound roxarsone." Analytical Methods 12, no. 47: 5729-5736.

Journal article
Published: 18 November 2020 in Sensors
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Multiplex lateral flow immunoassay (LFIA) is largely used for point-of-care testing to detect different pathogens or biomarkers in a single device. The increasing demand for multitargeting diagnostics requires multi-informative single tests. In this study, we demonstrated three strategies to upgrade standard multiplex LFIA to multimodal capacity. As a proof-of-concept, we applied the strategies to the differential diagnosis of Human Immunodeficiency Virus (HIV) infection, a widespread pathogen, for which conventional multiplex LFIA testing is well-established. In the new two-parameter LFIA (x2LFIA), we exploited color encoding, in which the binding of multiple targets occurs in one reactive band and the color of the probe reveals which one is present in the sample. By combining the sequential alignment of several reactive zones along the membrane of the LFIA strip and gold nanoparticles and gold nanostars for the differential visualization, in this demonstration, the x2LFIA can furnish information on HIV serotype and stage of infection in a single device. Three immunosensors were designed. The use of bioreagents as the capturing ligand anchored onto the membrane or as the detection ligand labelled with gold nanomaterials affected the performance of the x2LFIA. Higher detectability was achieved by the format involving the HIV-specific antigens as capturing agent and labelled secondary bioligands (anti-human immunoglobulins M and protein G) as the probes.

ACS Style

Simone Cavalera; Fabio Di Nardo; Luca Forte; Francesca Marinoni; Matteo Chiarello; Claudio Baggiani; Laura Anfossi. Switching from Multiplex to Multimodal Colorimetric Lateral Flow Immunosensor. Sensors 2020, 20, 6609 .

AMA Style

Simone Cavalera, Fabio Di Nardo, Luca Forte, Francesca Marinoni, Matteo Chiarello, Claudio Baggiani, Laura Anfossi. Switching from Multiplex to Multimodal Colorimetric Lateral Flow Immunosensor. Sensors. 2020; 20 (22):6609.

Chicago/Turabian Style

Simone Cavalera; Fabio Di Nardo; Luca Forte; Francesca Marinoni; Matteo Chiarello; Claudio Baggiani; Laura Anfossi. 2020. "Switching from Multiplex to Multimodal Colorimetric Lateral Flow Immunosensor." Sensors 20, no. 22: 6609.

Journal article
Published: 26 October 2020 in Biosensors and Bioelectronics
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To accurately diagnose COVID-19 infection and its time-dependent progression, the rapid, sensitive, and noninvasive determination of immunoglobulins A specific to SARS-CoV-2 (IgA) in saliva and serum is needed to complement tests that detect immunoglobulins G and M. We have developed a dual optical/chemiluminescence format of a lateral flow immunoassay (LFIA) immunosensor for IgA in serum and saliva. A recombinant nucleocapsid antigen specifically captures SARS-CoV-2 antibodies in patient specimens. A labelled anti-human IgA reveals the bound IgA fraction. A dual colorimetric and chemiluminescence detection enables the affordable and ultrasensitive determination of IgA to SARS-CoV-2. Specifically, a simple smartphone-camera-based device measures the colour signal provided by nanogold-labelled anti-human IgA. For the ultrasensitive chemiluminescence transduction, we used a contact imaging portable device based on cooled CCD, and measured the light signal resulting from the reaction of the HRP-labelled anti-human IgA with a H2O2/luminol/enhancers substrate. A total of 25 serum and 9 saliva samples from infected and/or recovered individuals were analysed by the colorimetric LFIA, which was sensitive and reproducible enough for the semi-quantification of IgA in subjects with a strong serological response and in the early stage of COVID-19 infection. Switching to CL detection, the same immunosensor exhibited higher detection capability, revealing the presence of salivary IgA in infected individuals. For the patients included in the study (n = 4), the level of salivary IgA correlated with the time elapsed from diagnosis and with the severity of the disease. This IgA-LFIA immunosensor could be useful for noninvasively monitoring early immune responses to COVID-19 and for investigating the diagnostic/prognostic utility of salivary IgA in the context of large-scale screening to assess the efficacy of SARS-CoV-2 vaccines.

ACS Style

Aldo Roda; Simone Cavalera; Fabio Di Nardo; Donato Calabria; Sergio Rosati; Patrizia Simoni; Barbara Colitti; Claudio Baggiani; Matilde Roda; Laura Anfossi. Dual lateral flow optical/chemiluminescence immunosensors for the rapid detection of salivary and serum IgA in patients with COVID-19 disease. Biosensors and Bioelectronics 2020, 172, 112765 -112765.

AMA Style

Aldo Roda, Simone Cavalera, Fabio Di Nardo, Donato Calabria, Sergio Rosati, Patrizia Simoni, Barbara Colitti, Claudio Baggiani, Matilde Roda, Laura Anfossi. Dual lateral flow optical/chemiluminescence immunosensors for the rapid detection of salivary and serum IgA in patients with COVID-19 disease. Biosensors and Bioelectronics. 2020; 172 ():112765-112765.

Chicago/Turabian Style

Aldo Roda; Simone Cavalera; Fabio Di Nardo; Donato Calabria; Sergio Rosati; Patrizia Simoni; Barbara Colitti; Claudio Baggiani; Matilde Roda; Laura Anfossi. 2020. "Dual lateral flow optical/chemiluminescence immunosensors for the rapid detection of salivary and serum IgA in patients with COVID-19 disease." Biosensors and Bioelectronics 172, no. : 112765-112765.

Paper
Published: 06 October 2020 in Journal of Materials Chemistry B
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High-affinity antibodies were generated to develop enzymatic and lateral flow immunoassays for monitoring tenofovir, a drug commonly used for treating HIV infection and used as a biomarker of adherence to the therapy.

ACS Style

Simone Cavalera; Consuelo Agulló; Josep Vicent Mercader; Fabio Di Nardo; Matteo Chiarello; Laura Anfossi; Claudio Baggiani; Antonio D'Avolio; Antonio Abad-Somovilla; Antonio Abad-Fuentes. Monoclonal antibodies with subnanomolar affinity to tenofovir for monitoring adherence to antiretroviral therapies: from hapten synthesis to prototype development. Journal of Materials Chemistry B 2020, 1 .

AMA Style

Simone Cavalera, Consuelo Agulló, Josep Vicent Mercader, Fabio Di Nardo, Matteo Chiarello, Laura Anfossi, Claudio Baggiani, Antonio D'Avolio, Antonio Abad-Somovilla, Antonio Abad-Fuentes. Monoclonal antibodies with subnanomolar affinity to tenofovir for monitoring adherence to antiretroviral therapies: from hapten synthesis to prototype development. Journal of Materials Chemistry B. 2020; ():1.

Chicago/Turabian Style

Simone Cavalera; Consuelo Agulló; Josep Vicent Mercader; Fabio Di Nardo; Matteo Chiarello; Laura Anfossi; Claudio Baggiani; Antonio D'Avolio; Antonio Abad-Somovilla; Antonio Abad-Fuentes. 2020. "Monoclonal antibodies with subnanomolar affinity to tenofovir for monitoring adherence to antiretroviral therapies: from hapten synthesis to prototype development." Journal of Materials Chemistry B , no. : 1.

Journal article
Published: 05 October 2020 in Talanta
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A rapid test for detecting total immunoglobulins directed towards the nucleocapsid protein (N) of severe acute syndrome coronavirus 2 (SARS CoV-2) was developed, based on a multi-target lateral flow immunoassay comprising two test lines. Both test lines bound to several classes of immunoglobulins (G, M, and A). Specific anti-SARS immunoglobulins were revealed by a colorimetric probe formed by N and gold nanoparticles. Targeting the total antibodies response to infection enabled achieving 100% diagnostic specificity (95.75–100, C.I. 95%, n = 85 healthy and with other infections individuals) and 94.6% sensitivity (84.9–98.9, C.I. 95%, n = 62 SARS CoV-2 infected subjects) as early as 7 days post confirmation of positivity. Agreeing results with a reference serological ELISA were achieved, except for the earlier detection capability of the rapid test. Follow up of the three seroconverting patients endorsed the hypothesis of the random rise of the different immunoglobulins and strengthened the ‘total antibodies’ approach for the trustworthy detection of serological response to SARS CoV-2 infection.

ACS Style

Simone Cavalera; Barbara Colitti; Sergio Rosati; Gianmarco Ferrara; Luigi Bertolotti; Chiara Nogarol; Cristina Guiotto; Celeste Cagnazzo; Marco Denina; Franca Fagioli; Fabio Di Nardo; Matteo Chiarello; Claudio Baggiani; Laura Anfossi. A multi-target lateral flow immunoassay enabling the specific and sensitive detection of total antibodies to SARS COV-2. Talanta 2020, 223, 121737 -121737.

AMA Style

Simone Cavalera, Barbara Colitti, Sergio Rosati, Gianmarco Ferrara, Luigi Bertolotti, Chiara Nogarol, Cristina Guiotto, Celeste Cagnazzo, Marco Denina, Franca Fagioli, Fabio Di Nardo, Matteo Chiarello, Claudio Baggiani, Laura Anfossi. A multi-target lateral flow immunoassay enabling the specific and sensitive detection of total antibodies to SARS COV-2. Talanta. 2020; 223 ():121737-121737.

Chicago/Turabian Style

Simone Cavalera; Barbara Colitti; Sergio Rosati; Gianmarco Ferrara; Luigi Bertolotti; Chiara Nogarol; Cristina Guiotto; Celeste Cagnazzo; Marco Denina; Franca Fagioli; Fabio Di Nardo; Matteo Chiarello; Claudio Baggiani; Laura Anfossi. 2020. "A multi-target lateral flow immunoassay enabling the specific and sensitive detection of total antibodies to SARS COV-2." Talanta 223, no. : 121737-121737.

Journal article
Published: 29 August 2020 in Sensors and Actuators B: Chemical
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This work describes the development and the application of a lateral flow biosensor for the detection of the prostate specific antigen in urine (uPSA). The biosensor allowed uPSA detection in 10 minutes with a limit of detection and a range of quantification respectively of 20 ng mL-1 and 37 – 420 ng mL-1, showing within and between-day coefficients of variation ≤ 13%. It showed 92% of accuracy and an almost perfect concordance with the reference electrochemiluminescence immunoassay method. The biosensor design provides the disappearance of the Test line signal at the cut-off concentration. This was achieved with a double layer sensing strategy, in which gold nanoparticles were functionalized with Staphylococcal protein A – a mediator – instead of anti-PSA antibody. This strategy allow making a fine-tune on the concentration of the specific antibody, obtaining an on/off switch of the Test line at the cut-off value. The cut-off value was also established in this work, based on the distribution of uPSA levels from 140 patients, who were suspected of prostate cancer and who underwent to first biopsy. The clinical application of the biosensor to predict repeat biopsy outcome in 28 patients showed sensitivity, specificity, positive and negative predictive values of 100%, 64%, 74% and 100%, respectively.

ACS Style

Fabio Di Nardo; Sergio Occhipinti; Paolo Gontero; Simone Cavalera; Matteo Chiarello; Claudio Baggiani; Laura Anfossi. Detection of urinary prostate specific antigen by a lateral flow biosensor predicting repeat prostate biopsy outcome. Sensors and Actuators B: Chemical 2020, 325, 128812 .

AMA Style

Fabio Di Nardo, Sergio Occhipinti, Paolo Gontero, Simone Cavalera, Matteo Chiarello, Claudio Baggiani, Laura Anfossi. Detection of urinary prostate specific antigen by a lateral flow biosensor predicting repeat prostate biopsy outcome. Sensors and Actuators B: Chemical. 2020; 325 ():128812.

Chicago/Turabian Style

Fabio Di Nardo; Sergio Occhipinti; Paolo Gontero; Simone Cavalera; Matteo Chiarello; Claudio Baggiani; Laura Anfossi. 2020. "Detection of urinary prostate specific antigen by a lateral flow biosensor predicting repeat prostate biopsy outcome." Sensors and Actuators B: Chemical 325, no. : 128812.

Journal article
Published: 21 May 2020 in Polymers
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It has been reported that in the molecular imprinting technique, the use of preformed oligomers instead of functional monomers increases the stability of the non-covalent interactions with the template molecule, providing a sharp gain in terms of binding properties for the resulting imprinted polymer. Based on this theory, we assumed that the delayed addition of template molecules to a polymerization mixture enhances the binding properties of the resulting polymer. To verify this hypothesis, we imprinted several mixtures of 4-vinylpyridine/ethylene dimethacrylate (1:6 mol/mol) in acetonitrile by adding diclofenac progressively later from the beginning of the polymerization process. After polymerization, the binding isotherms of imprinted and non-imprinted materials were measured in acetonitrile by partition equilibrium experiments. Binding data confirm our hypothesis, as imprinted polymers prepared by delayed addition, with delay times of 5 and 10 min, showed higher binding affinity (Keq = 1.37 × 104 L mol−1 and 1.80 × 104 L mol−1) than the polymer obtained in the presence of template at the beginning (Keq = 5.30 × 103 L mol−1). Similarly, an increase in the imprinting factor measured vs. the non-imprinted polymer in the binding selectivity with respect to mefenamic acid was observed. We believe that the delayed addition approach could be useful in prepar imprinted polymers with higher binding affinity and increased binding selectivity in cases of difficult imprinting polymerization.

ACS Style

Laura Anfossi; Simone Cavalera; Fabio Di Nardo; Giulia Spano; Cristina Giovannoli; Claudio Baggiani. Delayed Addition of Template Molecules Enhances the Binding Properties of Diclofenac-Imprinted Polymers. Polymers 2020, 12, 1178 .

AMA Style

Laura Anfossi, Simone Cavalera, Fabio Di Nardo, Giulia Spano, Cristina Giovannoli, Claudio Baggiani. Delayed Addition of Template Molecules Enhances the Binding Properties of Diclofenac-Imprinted Polymers. Polymers. 2020; 12 (5):1178.

Chicago/Turabian Style

Laura Anfossi; Simone Cavalera; Fabio Di Nardo; Giulia Spano; Cristina Giovannoli; Claudio Baggiani. 2020. "Delayed Addition of Template Molecules Enhances the Binding Properties of Diclofenac-Imprinted Polymers." Polymers 12, no. 5: 1178.

Journal article
Published: 20 April 2020 in Toxins
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The diffusion of the legalization of cannabis for recreational, medicinal and nutraceutical uses requires the development of adequate analytical methods to assure the safety and security of such products. In particular, aflatoxins are considered to pose a major risk for the health of cannabis consumers. Among analytical methods that allows for adequate monitoring of food safety, immunoassays play a major role thanks to their cost-effectiveness, high-throughput capacity, simplicity and limited requirement for equipment and skilled operators. Therefore, a rapid and sensitive enzyme immunoassay has been adapted to measure the most hazardous aflatoxin B1 in cannabis products. The assay was acceptably accurate (recovery rate: 78–136%), reproducible (intra- and inter-assay means coefficients of variation 11.8% and 13.8%, respectively), and sensitive (limit of detection and range of quantification: 0.35 ng mL−1 and 0.4–2 ng mL−1, respectively corresponding to 7 ng g−1 and 8–40 ng g−1 ng g−1 in the plant) and provided results which agreed with a HPLC-MS/MS method for the direct analysis of aflatoxin B1 in cannabis inflorescence and leaves. In addition, the carcinogenic aflatoxin B1 was detected in 50% of the cannabis products analyzed (14 samples collected from small retails) at levels exceeding those admitted by the European Union in commodities intended for direct human consumption, thus envisaging the need for effective surveillance of aflatoxin contamination in legal cannabis.

ACS Style

Fabio Di Nardo; Simone Cavalera; Claudio Baggiani; Matteo Chiarello; Marco Pazzi; Laura Anfossi. Enzyme Immunoassay for Measuring Aflatoxin B1 in Legal Cannabis. Toxins 2020, 12, 265 .

AMA Style

Fabio Di Nardo, Simone Cavalera, Claudio Baggiani, Matteo Chiarello, Marco Pazzi, Laura Anfossi. Enzyme Immunoassay for Measuring Aflatoxin B1 in Legal Cannabis. Toxins. 2020; 12 (4):265.

Chicago/Turabian Style

Fabio Di Nardo; Simone Cavalera; Claudio Baggiani; Matteo Chiarello; Marco Pazzi; Laura Anfossi. 2020. "Enzyme Immunoassay for Measuring Aflatoxin B1 in Legal Cannabis." Toxins 12, no. 4: 265.

Journal article
Published: 24 January 2020 in Analytica Chimica Acta
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A high affinity and selectivity DNA aptamer for aflatoxin B1 (AFB1) was designed through Genetic Algorithm (GA) based in silico maturation (ISM) strategy. The sequence of a known AFB1 aptamer (Patent: PCT/CA2010/001292, Apt1) applied as a probe in many aptasensors was modified using seven GA rounds to generate an initial library and three different generations of ss DNA oligonucleotides as new candidate aptamers. Molecular docking methodology was used to screen and analyze the best aptamer–AFB1 complexes. Also, a new pipeline was proposed to faithfully predict the tertiary structure of all single stranded DNA sequences. By the second generation, aptamer Apt1 sequence was optimized in the local search space and five aptamers including F20, g12, C52, C32 and H1 were identified as the best aptamers for AFB1. The selected aptamers were applied as probes in an unmodified gold nanoparticles-based aptasensor to evaluate their binding affinity to AFB1 and their selectivity against other mycotoxins (aflatoxins B2, G1, G2, M1, ochratoxin A and zearalenone). In addition, a novel direct fluorescent anisotropy aptamer assay was developed to confirm the binding interaction of the selected aptamers over AFB1. The ISM allowed the identification of an aptamer, F20, with up to 9.4 and 2 fold improvement in affinity and selectivity compared to the parent aptamer, respectively.

ACS Style

Maryam Mousivand; Laura Anfossi; Kowsar Bagherzadeh; Nadia Barbero; Amir Mirzadi-Gohari; Mohammad Javan-Nikkhah. In silico maturation of affinity and selectivity of DNA aptamers against aflatoxin B1 for biosensor development. Analytica Chimica Acta 2020, 1105, 178 -186.

AMA Style

Maryam Mousivand, Laura Anfossi, Kowsar Bagherzadeh, Nadia Barbero, Amir Mirzadi-Gohari, Mohammad Javan-Nikkhah. In silico maturation of affinity and selectivity of DNA aptamers against aflatoxin B1 for biosensor development. Analytica Chimica Acta. 2020; 1105 ():178-186.

Chicago/Turabian Style

Maryam Mousivand; Laura Anfossi; Kowsar Bagherzadeh; Nadia Barbero; Amir Mirzadi-Gohari; Mohammad Javan-Nikkhah. 2020. "In silico maturation of affinity and selectivity of DNA aptamers against aflatoxin B1 for biosensor development." Analytica Chimica Acta 1105, no. : 178-186.

Journal article
Published: 05 August 2019 in ACS Applied Materials & Interfaces
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Stable and efficient conjugates between antibodies and gold nanoparticles (GNP-Ab) are sought to develop highly sensitive and robust biosensors with applications in medicine, toxicology, food safety controls, and targeted drug delivery. Several strategies have been proposed for directing the antibody attachment to GNPs thus preserving antibody activity, including covalently coupling the antibody to a polymer grafted on GNP surface and exploiting the high affinity of bioreceptors as mediators for the binding. Both approaches also allow for shielding GNPs with a protective layer that guarantees the robustness of the conjugate. Notwithstanding, antibodies freely adsorb to GNP with high binding efficiency. The nonspecific adsorption is far more simple, fast, and inexpensive than any mediated coupling. Therefore, it is preferred for most applications, although it is considered to produce GNP-Ab with a limited activity. In this work, we compared three strategies for producing GNP-Ab, such as (i) covalent coupling mediated by a chemical layer, (ii) affinity-based binding mediated by a biomolecular layer composed of Staphylococcal protein A, and (iii) direct attachment via adsorption. The so-prepared GNP-Ab were employed as probes in a colorimetric lateral flow immunoassay (LFIA) for measuring salivary cortisol as a model biosensor that relies on the use of active GNP-Ab conjugates. Unexpectedly, the biosensors fabricated using the three probes were completely comparable in terms of their ability to measure salivary cortisol. Furthermore, we observed that the sensitivity of the LFIA primarily depended on the amount of the antibody bound to GNPs rather than on the method by which it was bound. The probes prepared using both the direct adsorption approach and mediated coupling via the biochemical mediator enabled development of point-of-care devices for the fast, sensitive, and reliable measurement of human salivary cortisol.

ACS Style

Fabio Di Nardo; Simone Cavalera; Claudio Baggiani; Cristina Giovannoli; Laura Anfossi. Direct vs Mediated Coupling of Antibodies to Gold Nanoparticles: The Case of Salivary Cortisol Detection by Lateral Flow Immunoassay. ACS Applied Materials & Interfaces 2019, 11, 32758 -32768.

AMA Style

Fabio Di Nardo, Simone Cavalera, Claudio Baggiani, Cristina Giovannoli, Laura Anfossi. Direct vs Mediated Coupling of Antibodies to Gold Nanoparticles: The Case of Salivary Cortisol Detection by Lateral Flow Immunoassay. ACS Applied Materials & Interfaces. 2019; 11 (36):32758-32768.

Chicago/Turabian Style

Fabio Di Nardo; Simone Cavalera; Claudio Baggiani; Cristina Giovannoli; Laura Anfossi. 2019. "Direct vs Mediated Coupling of Antibodies to Gold Nanoparticles: The Case of Salivary Cortisol Detection by Lateral Flow Immunoassay." ACS Applied Materials & Interfaces 11, no. 36: 32758-32768.

Research article
Published: 18 June 2019 in Nano Research
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Carbon nitride nanoparticles (CNNPs) have been employed as fluorescent sensing tools owing to their unique features, e.g. low cost production, high stability in water and high photoluminescence quantum yield. Here, an easy and versatile synthetic approach was exploited to design fluorescent nanoparticles with surface functionalities suitable for covalent binding to bioligands. High hydrophilic, brightly fluorescent CNNPs, rich of superficial amines, were obtained from the thermal condensation of urea and lysine (CNNPLys) and by tuning the precursor ratio and the heating time. Structure and size of the functionalized nanoparticles were characterized through infrared (IR) spectroscopy, transmission electron microscopy (TEM) and dynamic light scattering (DLS). Their optical properties were studied by ultraviolet-visible (UV-Vis) and fluorescence spectroscopy. The superficial primary amino groups, furnished by the lysine co-precursor, enabled for covalently linking CNNPLys to model proteins. The CNNPLys-protein conjugates excited under UV irradiation emit in the 400–450 nm visible range (quantum yield 24%). The applicability of CNNPLys as novel fluorescent probes was demonstrated by a fluorescence quenching assay, in which gold nanoparticles (GNPs) were attached to Staphylococcal protein A and employed to quench the CNNPLys fluorescence by resonant energy transfer (FRET). The quenching occurred upon formation of the specific binding between the GNP-linked protein A and CNNPLys-tagged immunoglobulins, while the inhibition of the binding resulted in the recovery of CNNPLys luminescence. The synthetic strategy, based on combining a “conjugated polymer”-forming unit (urea) and a co-precursor able to provide the desired functional group (lysine), allows designing innovative materials for the development of new generation fluorescence biosensors in which easily functionalized fluorophores are needed.

ACS Style

Gabriele Capilli; Simone Cavalera; Laura Anfossi; Cristina Giovannoli; Marco Minella; Claudio Baggiani; Claudio Minero. Amine-rich carbon nitride nanoparticles: Synthesis, covalent functionalization with proteins and application in a fluorescence quenching assay. Nano Research 2019, 12, 1862 -1870.

AMA Style

Gabriele Capilli, Simone Cavalera, Laura Anfossi, Cristina Giovannoli, Marco Minella, Claudio Baggiani, Claudio Minero. Amine-rich carbon nitride nanoparticles: Synthesis, covalent functionalization with proteins and application in a fluorescence quenching assay. Nano Research. 2019; 12 (8):1862-1870.

Chicago/Turabian Style

Gabriele Capilli; Simone Cavalera; Laura Anfossi; Cristina Giovannoli; Marco Minella; Claudio Baggiani; Claudio Minero. 2019. "Amine-rich carbon nitride nanoparticles: Synthesis, covalent functionalization with proteins and application in a fluorescence quenching assay." Nano Research 12, no. 8: 1862-1870.

Paper
Published: 25 March 2019 in Analytical Methods
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A molecularly imprinted sorbent assays for cortisol was optimized for direct determination in human saliva.

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Giulia Spano; Simone Cavalera; Fabio Di Nardo; Cristina Giovannoli; Laura Anfossi; Claudio Baggiani. Development of a biomimetic enzyme-linked immunosorbent assay based on a molecularly imprinted polymer for the detection of cortisol in human saliva. Analytical Methods 2019, 11, 2320 -2326.

AMA Style

Giulia Spano, Simone Cavalera, Fabio Di Nardo, Cristina Giovannoli, Laura Anfossi, Claudio Baggiani. Development of a biomimetic enzyme-linked immunosorbent assay based on a molecularly imprinted polymer for the detection of cortisol in human saliva. Analytical Methods. 2019; 11 (17):2320-2326.

Chicago/Turabian Style

Giulia Spano; Simone Cavalera; Fabio Di Nardo; Cristina Giovannoli; Laura Anfossi; Claudio Baggiani. 2019. "Development of a biomimetic enzyme-linked immunosorbent assay based on a molecularly imprinted polymer for the detection of cortisol in human saliva." Analytical Methods 11, no. 17: 2320-2326.

Review
Published: 26 December 2018 in Biosensors
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Simultaneous measurement of different substances from a single sample is an emerging issue for achieving efficient and high-throughput detection in several fields of application. Although immunoanalytical techniques have well-established and prevailing advantages over alternative screening analytical platforms, one of the incoming challenges for immunoassay is exact multiplexing. Lateral flow immunoassay (LFIA) is a leading immunoanalytical technique for onsite analysis, thanks to its simplicity, rapidity, and cost-effectiveness. Moreover, LFIA architecture is adaptable to multiplexing, and is therefore a possible answer to the pressing demand of multiplexing point-of-need analysis. This review presents an overview of diverse approaches for multiplex LFIA, with a special focus on strategies based on new types of magnetic, fluorescent, and colored labels.

ACS Style

Laura Anfossi; Fabio Di Nardo; Simone Cavalera; Cristina Giovannoli; Claudio Baggiani. Multiplex Lateral Flow Immunoassay: An Overview of Strategies towards High-throughput Point-of-Need Testing. Biosensors 2018, 9, 2 .

AMA Style

Laura Anfossi, Fabio Di Nardo, Simone Cavalera, Cristina Giovannoli, Claudio Baggiani. Multiplex Lateral Flow Immunoassay: An Overview of Strategies towards High-throughput Point-of-Need Testing. Biosensors. 2018; 9 (1):2.

Chicago/Turabian Style

Laura Anfossi; Fabio Di Nardo; Simone Cavalera; Cristina Giovannoli; Claudio Baggiani. 2018. "Multiplex Lateral Flow Immunoassay: An Overview of Strategies towards High-throughput Point-of-Need Testing." Biosensors 9, no. 1: 2.

Research paper
Published: 06 November 2018 in Analytical and Bioanalytical Chemistry
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In this study, we report the simultaneous use of gold and silver nanoparticles to set a multicolor multiplex lateral flow immunoassay (xLFIA). Silver nanoparticles (AgNPs), spherical in shape and characterized by a brilliant yellow color, were obtained by a new viable one-step synthetic protocol. AgNPs were stable over time and acceptably robust to conditions used for fabricating LFIA devices. These AgNPs were employed as a colorimetric probe in combination with two different kinds of gold nanoparticles (AuNPs) to set a visual xLFIA for detecting allergens. Surface plasmon resonance peaks of probes (AgNPs, spherical and desert rose-like AuNPs) were centered at 420, 525, and 620 nm, respectively. Therefore, the xLFIA output was easily interpreted through a “yellow magenta cyan (YMC)” color code. The prospect of the YMC xLFIA was demonstrated by simultaneously detecting three major allergens in bakery products. Antibodies directed towards casein, ovalbumin, and hazelnut allergenic proteins were individually adsorbed onto metal nanoparticles to produce three differently colored specific probes. These were inserted in a LFIA comprising three lines, each responsive for one allergen. The trichromatic xLFIA was able to detect allergenic proteins at levels as low as 0.1 mg/l and enabled the easy identification of the allergens in commercial biscuits based on the color of the probes.

ACS Style

Laura Anfossi; Fabio Di Nardo; Alida Russo; Simone Cavalera; Cristina Giovannoli; Giulia Spano; Sabine Baumgartner; Kathrin Lauter; Claudio Baggiani. Silver and gold nanoparticles as multi-chromatic lateral flow assay probes for the detection of food allergens. Analytical and Bioanalytical Chemistry 2018, 411, 1905 -1913.

AMA Style

Laura Anfossi, Fabio Di Nardo, Alida Russo, Simone Cavalera, Cristina Giovannoli, Giulia Spano, Sabine Baumgartner, Kathrin Lauter, Claudio Baggiani. Silver and gold nanoparticles as multi-chromatic lateral flow assay probes for the detection of food allergens. Analytical and Bioanalytical Chemistry. 2018; 411 (9):1905-1913.

Chicago/Turabian Style

Laura Anfossi; Fabio Di Nardo; Alida Russo; Simone Cavalera; Cristina Giovannoli; Giulia Spano; Sabine Baumgartner; Kathrin Lauter; Claudio Baggiani. 2018. "Silver and gold nanoparticles as multi-chromatic lateral flow assay probes for the detection of food allergens." Analytical and Bioanalytical Chemistry 411, no. 9: 1905-1913.

Book chapter
Published: 26 September 2018 in Rapid Test - Advances in Design, Format and Diagnostic Applications
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Laura Anfossi; Cristina Giovannoli; Claudio Baggiani. Introductory Chapter: Rapid Test - Advances in Design, Formats, and Detection Strategies. Rapid Test - Advances in Design, Format and Diagnostic Applications 2018, 1 .

AMA Style

Laura Anfossi, Cristina Giovannoli, Claudio Baggiani. Introductory Chapter: Rapid Test - Advances in Design, Formats, and Detection Strategies. Rapid Test - Advances in Design, Format and Diagnostic Applications. 2018; ():1.

Chicago/Turabian Style

Laura Anfossi; Cristina Giovannoli; Claudio Baggiani. 2018. "Introductory Chapter: Rapid Test - Advances in Design, Formats, and Detection Strategies." Rapid Test - Advances in Design, Format and Diagnostic Applications , no. : 1.

Journal article
Published: 18 September 2018 in Talanta
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A multiplex Lateral Flow Immunoassay was developed based on the use of a single Test line and multicolour gold nanoparticles (GNPs) as signal reporters. Red and blue GNPs were linked to antibodies directed towards two different analytes and included in a typical lateral flow immunoassay configuration, in which the Test line was formed by the mixture of two antigens. As a result of the immunoreactions occurring at the Test zone, diverse combinations of red and blue GNPs labels were captured. Therefore, the Test line assumed different colours depending on which – and how much – analyte is present in the sample. The multiplexing capability of the ‘colour-encoded assay’ is illustrated by the simultaneous detection of aflatoxin B1 (AFB1) and type-B fumonisins (FMs) in wheat and food products that made with wheat. Reproducible detection of AFB1 and FMs contamination in raw and processed food was achieved with visual cut-off levels at 1 ng mL−1 and 50 ng mL−1, respectively. The contaminant was identified based on the colour of the label according with a specific colour code. Furthermore, strips images were acquired by means of a common smartphone and analysed through RGB data analysis providing semi-quantitative detection of the two mycotoxins.

ACS Style

Fabio Di Nardo; Eugenio Alladio; Claudio Baggiani; Simone Cavalera; Cristina Giovannoli; Giulia Spano; Laura Anfossi. Colour-encoded lateral flow immunoassay for the simultaneous detection of aflatoxin B1 and type-B fumonisins in a single Test line. Talanta 2018, 192, 288 -294.

AMA Style

Fabio Di Nardo, Eugenio Alladio, Claudio Baggiani, Simone Cavalera, Cristina Giovannoli, Giulia Spano, Laura Anfossi. Colour-encoded lateral flow immunoassay for the simultaneous detection of aflatoxin B1 and type-B fumonisins in a single Test line. Talanta. 2018; 192 ():288-294.

Chicago/Turabian Style

Fabio Di Nardo; Eugenio Alladio; Claudio Baggiani; Simone Cavalera; Cristina Giovannoli; Giulia Spano; Laura Anfossi. 2018. "Colour-encoded lateral flow immunoassay for the simultaneous detection of aflatoxin B1 and type-B fumonisins in a single Test line." Talanta 192, no. : 288-294.

Evaluation study
Published: 17 September 2018 in Biosensors and Bioelectronics
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During space missions, real-time monitoring of astronauts’ health status is of crucial importance and therefore there is a strong demand for simple analytical devices that astronauts can use to perform clinical chemistry analyses directly onboard. As part of the “IN SITU Bioanalysis” project, we designed a biosensor for analyzing salivary levels of cortisol in astronauts, a marker of chronic stress. The biosensor is based on the Lateral Flow Immunoassay (LFIA) approach coupled with chemiluminescence (CL) detection and comprised a 3D-printed plastic cartridge containing a sealed fluidic element with the LFIA strip, in which the flow of sample and reagents was activated by pressing buttons on the cartridge and sustained by exploiting capillary forces. For measurement, the photon emission was imaged employing a CL reader based on an ultrasensitive cooled charge-coupled device (CCD) camera. The payload was designed to operate in microgravity and to withstand mechanical stress, such as take-off vibrations, and onboard depressurization events, while the microfluidics was developed considering alterations of physical phenomena occurring in microgravity, such as bubble formation, surface wettability and liquid evaporation. The biosensor, which was successfully used by the Italian astronaut Paolo Nespoli during the VITA mission (July-December 2017), demonstrated the feasibility of performing sensitive LFIA analysis of salivary cortisol down to 0.4 ng/mL directly onboard the International Space Station. It could be easily adapted for the analysis of other clinical biomarkers, thus enabling the early diagnosis of diseases and the timely activation of appropriate countermeasures.

ACS Style

Martina Zangheri; Mara Mirasoli; Massimo Guardigli; Fabio Di Nardo; Laura Anfossi; Claudio Baggiani; Patrizia Simoni; Mario Benassai; Aldo Roda. Chemiluminescence-based biosensor for monitoring astronauts’ health status during space missions: Results from the International Space Station. Biosensors and Bioelectronics 2018, 129, 260 -268.

AMA Style

Martina Zangheri, Mara Mirasoli, Massimo Guardigli, Fabio Di Nardo, Laura Anfossi, Claudio Baggiani, Patrizia Simoni, Mario Benassai, Aldo Roda. Chemiluminescence-based biosensor for monitoring astronauts’ health status during space missions: Results from the International Space Station. Biosensors and Bioelectronics. 2018; 129 ():260-268.

Chicago/Turabian Style

Martina Zangheri; Mara Mirasoli; Massimo Guardigli; Fabio Di Nardo; Laura Anfossi; Claudio Baggiani; Patrizia Simoni; Mario Benassai; Aldo Roda. 2018. "Chemiluminescence-based biosensor for monitoring astronauts’ health status during space missions: Results from the International Space Station." Biosensors and Bioelectronics 129, no. : 260-268.