Re-modeling ELISA kits embedded in an automated system suitable for on-line detection of algal toxins in seawater

Πετρόπουλος, Κωνσταντίνος; Bodini, Sergio F.; Fabiani, Laura; Micheli, Laura; Porchetta, Alessandro; Piermarini, S.; Volpe, G.; Pasquazzi, Fabio Massimo; Sanfilippo, Luca; Moscetta, Pompeo; Chiavarini, Salvatore; Palleschi, Giuseppe

doi:10.1016/j.snb.2018.12.083

Cited by 30 publications

(7 citation statements)

References 33 publications

(35 reference statements)

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“…1 The automatic instruments, standard work-ow and commercial reagent kit enable ELISA to be widely used in clinical testing and biological research, 2 playing a more and more important role in disease screening, 3,4 food safety 5,6 and environmental monitoring. 7,8 In clinical test, ELISA usually adopts uorescence or luminescence as detection signal. 9 The uorescence intensity is decreased with the target protein concentration, which causes ELISA is not able to discover the abnormal expression of proteins at the early stage of diseases.…”

Section: Introductionmentioning

confidence: 99%

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Ball-lens assisted sensitivity improvement of fluorescence immunoassay in microchannels

Zhang

et al. 2021

RSC Adv.

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Section: Introductionmentioning

confidence: 99%

“… 1 The automatic instruments, standard work-flow and commercial reagent kit enable ELISA to be widely used in clinical testing and biological research, 2 playing a more and more important role in disease screening, 3,4 food safety 5,6 and environmental monitoring. 7,8 …”

Section: Introductionmentioning

confidence: 99%

Ball-lens assisted sensitivity improvement of fluorescence immunoassay in microchannels

Zhang

et al. 2021

RSC Adv.

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“…Traditional approaches for STX analysis include mouse bioassay [ 8 ], enzyme-linked immunosorbent assay (ELISA) [ 9 ], cell assay [ 10 ], high-performance liquid chromatography (HPLC) [ 11 ], and liquid chromatography-tandem mass spectrometry (LC-MS) [ 12 ]. Although respective advantages in sensitivity and specificity have been achieved by these methods, there are some challenges: for example, HPLC and LCMS need sophisticated equipment and complex pre-treatments [ 13 , 14 ], and ELISA requires tedious lab work [ 15 ]. To address these challenges, electrochemical analysis with functional nanomaterials could be a potential solution [ 16 ].…”

Section: Introductionmentioning

confidence: 99%

An Electrochemical Ti3C2Tx Aptasensor for Sensitive and Label-Free Detection of Marine Biological Toxins

Ullah

Chen

Noureen

et al. 2021

Sensors

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Saxitoxin (STX) belongs to the family of marine biological toxins, which are major contaminants in seafood. The reference methods for STX detection are mouse bioassay and chromatographic analysis, which are time-consuming, high costs, and requirement of sophisticated operation. Therefore, the development of alternative methods for STX analysis is urgent. Electrochemical analysis is a fast, low-cost, and sensitive method for biomolecules analysis. Thus, in this study, an electrolyte-insulator-semiconductor (EIS) sensor based on aptamer-modified two-dimensional layered Ti3C2Tx nanosheets was developed for STX detection. The high surface area and rich functional groups of MXene benefited the modification of aptamer, which had specific interactions with STX. Capacitance-voltage (C-V) and constant-capacitance (ConCap) measurement results indicated that the aptasensor was able to detect STX with high sensitivity and good specificity. The detection range was 1.0 nM to 200 nM and detection limit was as low as 0.03 nM. Moreover, the aptasensor was found to have a good selectivity and two-week stability. The mussel tissue extraction test suggested the potential application of this biosensor in detecting STX in real samples. This method provides a convenient approach for low-cost, rapid, and label-free detection of marine biological toxins.

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“…This overcrowding can give rise to a phenomenon of accumulation in seawater animals (mollusks, shellfish, and fishes), causing intoxication along the entire food chain that involves cetaceans, birds, other mammals, and finally humans. Over the past few years, many different analytical approaches have been explored in the area of algal toxin detection from environmental samples, including liquid chromatography coupled with mass-spectroscopy (LC-MS) ( Vilariño et al, 2013 ), high-performance liquid chromatography (HPLC) ( Van Egmond et al, 1994 ), enzyme-linked immunosorbent assay (ELISA) ( Micheli et al, 2002 ; Yu et al, 2002 ; Petropoulos et al, 2019 ), and electrochemical biosensors ( Zhang et al, 2018 ). Most of the research has been focused on the saxitoxin (STX) and STX-related compounds, as they are found to be more common in neurotoxic paralytic shellfish toxins (PSTs) ( Deeds et al, 2008 ).…”

Section: Introductionmentioning

confidence: 99%

Microfluidic Flow Injection Immunoassay System for Algal Toxins Determination: A Case of Study

et al. 2021

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A novel flow injection microfluidic immunoassay system for continuous monitoring of saxitoxin, a lethal biotoxin, in seawater samples is presented in this article. The system consists of a preimmobilized G protein immunoaffinity column connected in line with a lab-on-chip setup. The detection of saxitoxin in seawater was carried out in two steps: an offline incubation step (competition reaction) performed between the analyte of interest (saxitoxin or Ag, as standard or seawater sample) and a tracer (an enzyme-conjugated antigen or Ag*) toward a specific polyclonal antibody. Then, the mixture was injected through a “loop” of a few μL using a six-way injection valve into a bioreactor, in line with the valve. The bioreactor consisted of a small glass column, manually filled with resin upon which G protein has been immobilized. When the mixture flowed through the bioreactor, all the antibody-antigen complex, formed during the competition step, is retained by the G protein. The tracer molecules that do not interact with the capture antibody and protein G are eluted out of the column, collected, and mixed with an enzymatic substrate directly within the microfluidic chip, via the use of two peristaltic pumps. When Ag* was present, a color change (absorbance variation, ΔAbs) of the solution is detected at a fixed wavelength (655 nm) by an optical chip docking system and registered by a computer. The amount of saxitoxin, present in the sample (or standard), that generates the variation of the intensity of the color, will be directly proportional to the concentration of the analyte in the analyzed solution. Indeed, the absorbance response increased proportionally to the enzymatic product and to the concentration of saxitoxin in the range of 3.5 × 10–7–2 × 10–5 ng ml−1 with a detection limit of 1 × 10–7 ng ml−1 (RSD% 15, S N−1 equal to 3). The immunoanalytical system has been characterized, optimized, and tested with seawater samples. This analytical approach, combined with the transportable and small-sized instrumentation, allows for easy in situ monitoring of marine water contaminations.

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Re-modeling ELISA kits embedded in an automated system suitable for on-line detection of algal toxins in seawater

Cited by 30 publications

References 33 publications

Ball-lens assisted sensitivity improvement of fluorescence immunoassay in microchannels

Ball-lens assisted sensitivity improvement of fluorescence immunoassay in microchannels

An Electrochemical Ti3C2Tx Aptasensor for Sensitive and Label-Free Detection of Marine Biological Toxins

Microfluidic Flow Injection Immunoassay System for Algal Toxins Determination: A Case of Study

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