Thermometric enzyme linked immunosorbent assay in continuous flow system: Optimization and evaluation using human serum albumin as a model system

Borrebaeck, Carl; Börjeson, Jan; Mattìasson, Bo

doi:10.1016/0009-8981(78)90381-9

Cited by 25 publications

(4 citation statements)

References 18 publications

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“…The results showed good between and within-column repeatability. As discussed earlier in reusability, the calibration with B 0 improves the reliability of the repeated analyses [35].…”

Section: Precisionmentioning

confidence: 81%

Development of an automated flow‐based spectrophotometric immunoassay for continuous detection of zearalenone

Jantra

Zór

Sanders

et al. 2020

Biotechnology and Applied Biochemistry

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“…The results showed good between and within-column repeatability. As discussed earlier in reusability, the calibration with B 0 improves the reliability of the repeated analyses [35].…”

Section: Precisionmentioning

confidence: 81%

Development of an automated flow‐based spectrophotometric immunoassay for continuous detection of zearalenone

Jantra

Zór

Sanders

et al. 2020

Biotechnology and Applied Biochemistry

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“…Blue line (triangle) shows number of assay cycles run. Grey line (square) shows the peak height in percent of a preceding pure aggregate pulse obtained for reference samples containing 40 µg HSA/mL sample (Modified from [3]).…”

Section: Figurementioning

confidence: 99%

“…Then, the number of antibody binding sites was limiting, and denaturation then resulted in decreased sensitivity of the assay. One can compensate for these negative effects by measuring the binding of a standardized samples of labeled antigen [3]. If this is taken as 100% signal, then assay of the native antigen in the competitive assay will stay constant, as seen in Figure 1.…”

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confidence: 99%

Why Using Molecularly Imprinted Polymers in Connection to Biosensors?

Mattìasson

Ertürk

2017

Sensors

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The area of biosensor-oriented research has grown rapidly during recent years. From a start with enzyme-based sensors, it soon moved over to affinity-based sensors, e.g., immuno-based sensors. As long as the analytical device was used for a single measurement, i.e., as a disposable, stability problems were connected to storage stability, but not to operational stability. However, when repeated assays were to be used, or even sensors for on-line monitoring, then operational stability also came into play [1].To secure high operational stability, one often operates in enzyme technology studies with a surplus of enzymes. That means that the process is diffusion-controlled, and when enzyme molecules denature and lose activity, then earlier resting enzymes come into action [2].For immuno-based assays, one can approach the stability problems in two ways. The initial studies were made with competitive binding assays using labeled antigens mixed with free antigens. Then, the number of antibody binding sites was limiting, and denaturation then resulted in decreased sensitivity of the assay. One can compensate for these negative effects by measuring the binding of a standardized samples of labeled antigen [3]. If this is taken as 100% signal, then assay of the native antigen in the competitive assay will stay constant, as seen in Figure 1. For direct binding assays, an excess of binding sites is needed, and denaturation of a fraction thus does not influence sensitivity since earlier resting antibodies come into play instead of those that denatured.Most binding assays involving MIPs are at present direct binding assays; thus, one shall use a surplus of the binding sites.There are many reasons why a sensor with an immobilized biomolecule (enzyme, antibody, etc.) is losing activity. Denaturation due to temperature effects is often regarded as a causative; therefore, a refrigerator is recommended for storage of the bioactive components. The low temperature tolerance restricts the use of enzyme-based sensors in, e.g., fermentation control. It has so far, in essence, been impossible to operate with enzyme-based biosensors directly in fermentation broth. Off-line analysis has been the alternative choice. Thermostable enzymes from extremophilic microorganisms may help to some extent, but not enough for standing heat sterilization.In areas where an unbroken cold-chain cannot be guaranteed, then biosensors based on selectivity of biological macromolecules constitute a problem. That may be a problem in developing countries, esp. when analysis is planned in remote areas.However, there are other problems that might appear, e.g., the presence of proteases in the medium to be analyzed (that was what happened in the experiment illustrated in Figure 1). Such enzymes may destroy the bioactive component on the sensor surface, thereby reducing the sensitivity and definitely destroying the reproducibility. There are examples when infections of microorganisms have led to the presence of proteases that quickly destroyed the sensor device (Figure...

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“…In dcTELISA, 7 the analyte and a constant amount of labeled derivative compete for the binding sites on the Ab, which are covalently immobilized onto CPG. Aer competition, the immunocomplexes form on an immunosorbent packed in a reactor.…”

Section: Introductionmentioning

confidence: 99%

Fast detection of atrazine in corn using thermometric biosensors

Qie¹,

Ning²,

Liu³

et al. 2013

Analyst

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Fast detection is important in screening large-scale samples. This study establishes a direct competitive ELISA method (dcTELISA) based on an enzyme thermistor for fast atrazine (ATZ) detection. ATZ competes with β-lactamase-labeled ATZ (ATZ-E) for the binding sites on anti-ATZ monoclonal antibody (mAb). The mAb are covalently bound to Controlled Pore Glass (CPG) in an immunoreactor to form immunocomplexes with ATZ and ATZ-E. Several parameters of biosensor performance were optimized, such as the ATZ-E concentration, concentration and nature of the substrate, flow rate, and effect of temperature on the sensor response. After optimization, the assay time for a single sample was 12 min. The work process and result were compared with those of high-performance liquid chromatography (HPLC). The detection results exhibited a recovery rate of 88% to 107% in ATZ-spiked fresh cut corn stalks and silage samples. The results obtained via dcTELISA had good correlation with that of HPLC, and the biosensor response was reproducible and stable even when used continuously for over 4 months. All these properties suggested that the fast detection method, dcTELISA, may be used to detect pesticide residue in large-scale samples.

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Thermometric enzyme linked immunosorbent assay in continuous flow system: Optimization and evaluation using human serum albumin as a model system

Cited by 25 publications

References 18 publications

Development of an automated flow‐based spectrophotometric immunoassay for continuous detection of zearalenone

Development of an automated flow‐based spectrophotometric immunoassay for continuous detection of zearalenone

Why Using Molecularly Imprinted Polymers in Connection to Biosensors?

Fast detection of atrazine in corn using thermometric biosensors

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