Preparation of Monoclonal Antibody for Brevetoxin 1 and Development of Ic-ELISA and Colloidal Gold Strip to Detect Brevetoxin 1

Ling, Sumei; Xiao, Shiwei; Xie, Cheng-Jie; Wang, Rongzhi; Zeng, Linmao; Wang, Ke; Zhang, Danping; Li, Xiulan; Wang, Shihua

doi:10.3390/toxins10020075

Cited by 22 publications

(12 citation statements)

References 24 publications

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“…Among detection approaches, including enzyme-linked immunosorbent assay (ELISA), colloidal gold immunochromatographic strips have the advantages of simple operation, strong specificity and high sensitivity, does not need expensive instruments and equipment and professional operators, and is very suitable for on-site rapid detection and screening of actual samples, and some commercial ELISA kits have been put into practical application. However, the stability of antibodies is relatively low, the preparation of antibodies is tedious, time-consuming, and expensive [23,24]. Especially for small molecular substances such as marine toxins, the preparation of antibody is more difficult because of defects of their low immunogenicity, high toxicity, and cross-reaction [23-25].…”

Section: The Classical Detection Methods For Marine Toxinsmentioning

confidence: 99%

“…In recent years, with the aggravation of environmental pollution, human deaths due to accidental ingestion of toxic shellfish are also often reported [18][19][20][21]. At present, the main methods for the detection of marine toxins include mouse bioassay (MBA) [22], enzyme-linked immunosorbent assay (ELISA) [23][24][25], high-performance liquid chromatography (HPLC) and liquid chromatography-mass spectrometry (LC-MS/MS), etc. [26][27][28].…”

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confidence: 99%

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Marine Toxins Detection by Biosensors Based on Aptamers

Wei

Liu

Zhang

et al. 2019

Toxins

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Marine toxins cause great harm to human health through seafood, therefore, it is urgent to exploit new marine toxins detection methods with the merits of high sensitivity and specificity, low detection limit, convenience, and high efficiency. Aptasensors have emerged to replace classical detection methods for marine toxins detection. The rapid development of molecular biological approaches, sequencing technology, material science, electronics and chemical science boost the preparation and application of aptasensors. Taken together, the aptamer-based biosensors would be the best candidate for detection of the marine toxins with the merits of high sensitivity and specificity, convenience, time-saving, relatively low cost, extremely low detection limit, and high throughput, which have reduced the detection limit of marine toxins from nM to fM. This article reviews the detection of marine toxins by aptamer-based biosensors, as well as the selection approach for the systematic evolution of ligands by exponential enrichment (SELEX), the aptamer sequences. Moreover, the newest aptasensors and the future prospective are also discussed, which would provide thereotical basis for the future development of marine toxins detection by aptasensors.Key Contribution: This article reviews systematically the toxicity of marine toxins, the development of aptarmer-based biosensors, and progress in the marine toxin detection by aptasensors.Toxins 2020, 12, 1 2 of 22 selection technology and biosensor fabrication technology offer a new solution for the detection of marine toxins with high efficiency and sensitivity [13][14][15][16][17].Marine toxins are a class of small molecular compounds mainly produced by red tide algae. In recent years, with the aggravation of environmental pollution, human deaths due to accidental ingestion of toxic shellfish are also often reported [18][19][20][21]. At present, the main methods for the detection of marine toxins include mouse bioassay (MBA) [22], enzyme-linked immunosorbent assay (ELISA) [23][24][25], high-performance liquid chromatography (HPLC) and liquid chromatography-mass spectrometry (LC-MS/MS), etc. [26][27][28]. However, these traditional methods have some drawbacks including the requirement of a technician, poor repeatability, expensive equipment, and issues related to animal ethics [22]. As a new detection technology, the biosensor is a kind of analysis systems using cell molecules and other bio-materials as sensitive elements, combined with a secondary sensor to detect a variety of chemicals by cascade amplified signal [29]. Because of their advantages of simple operation, high speed, high sensitivity, miniaturization, and easy automation, biosensors have been widely used in different fields including cell physiology [30], drug screening [31,32], food safety detection [33], disease diagnosis [34], etc.Aptamer, meaning "to fit", is a kind of oligonucleotide which can bind to target molecules with high selectivity and affinity. On account of the existence of aptamers in nature and the po...

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Section: The Classical Detection Methods For Marine Toxinsmentioning

confidence: 99%

mentioning

confidence: 99%

Marine Toxins Detection by Biosensors Based on Aptamers

Wei

Liu

Zhang

et al. 2019

Toxins

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“…To the best of our knowledge, the success rate of hybridoma cell construction may be affected by many factors, such as the growth state of the myeloma cells, the concentration of PEG, the ratio of spleen cells to myeloma cells, and the number of feeder layer cells (35). In the present study, 7% PEG was used for cell fusion at 1 min of incubation.…”

Section: Discussionmentioning

confidence: 92%

Biological characterisation and application of human MTH1 and monoclonal antibody preparation

Chen

Li²,

et al. 2018

Oncol Rep

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Human MutT homolog 1 (MTH1) hydrolyses oxidised nucleotide triphosphates, thereby preventing them from being incorporated into DNA; MTH1 has been found to be elevated in many types of cancers, including lung, stomach cancer, melanoma and breast cancer. Thus, tumour-targeted hMTH1 may be valuable for developing novel anticancer therapies. In the present study, we prepared human MTH1 protein and its monoclonal antibody (mAb). The hMTH1 gene was cloned into the prokaryotic expression vector pET28a and optimally expressed in the E. coli Transetta (DE3) strain. Using an Ni-NTA column and a G-50 gel filtration column, 20.1 mg of active hMTH1 was obtained from 1,000 ml of bacterial culture, and the purity was over 98%, as detected by high-performance liquid chromatography (HPLC). The half maximal inhibitory concentration (IC 50 ) of TH287 (hMTH1 inhibitor) was determined to be 3.53±0.47 nM using the recombinant hMTH1 protein (rhMTH1). The enzyme activity assay showed the Michaelis constant (K m ) and the catalytic constant (k cat ) of the protein were 106.13±48.83 µM and 3.64±0.58 sec -1 , respectively. The anti-hMTH1 mAb was obtained via the hybridoma technique and validated by western blot analysis. In addition, an immunofluorescence assay (IFA) and ELISA determined that the mAb could efficiently bind to natural hMTH1 expressed on the human breast cancer cell line MCF-7. Taken together, the results showed the rhMTH1 is an active protein and has practical applications for inhibitor selection, and our prepared hMTH1 mAb will provide a valuable tool for the further characterisation of hMTH1 and antitumour medicinal development in future.

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“…Ling et al developed an ELISA capable of detecting BTX at levels as low as 14 ng/mL [ 115 ]. This assay was tested with a wide range of artificially spiked shellfish samples, including clams, mussels, oysters, and scallops and found to have an average toxin recovery rate of 89 ± 2%.…”

Section: Methods For End-product Testing (Ept)mentioning

confidence: 99%

“…Uniquely, Shen et al used quantum dots and gold nanoflowers to create an LFIA that could detect TTX concentrations as low as 0.2 ng/mL [ 132 ]. Ling et al adapted their BTX ELISA (discussed above) into an LFIA, however, unfortunately, the new LFIA format had significantly reduced sensitivity (200 ng/mL) compared to the ELISA format (14 ng/mL) [ 115 ].…”

Section: Methods For End-product Testing (Ept)mentioning

confidence: 99%

Current Trends and Challenges for Rapid SMART Diagnostics at Point-of-Site Testing for Marine Toxins

Dillon

Zaczek-Moczydlowska

Edwards

et al. 2021

Sensors

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In the past twenty years marine biotoxin analysis in routine regulatory monitoring has advanced significantly in Europe (EU) and other regions from the use of the mouse bioassay (MBA) towards the high-end analytical techniques such as high-performance liquid chromatography (HPLC) with tandem mass spectrometry (MS). Previously, acceptance of these advanced methods, in progressing away from the MBA, was hindered by a lack of commercial certified analytical standards for method development and validation. This has now been addressed whereby the availability of a wide range of analytical standards from several companies in the EU, North America and Asia has enhanced the development and validation of methods to the required regulatory standards. However, the cost of the high-end analytical equipment, lengthy procedures and the need for qualified personnel to perform analysis can still be a challenge for routine monitoring laboratories. In developing regions, aquaculture production is increasing and alternative inexpensive Sensitive, Measurable, Accurate and Real-Time (SMART) rapid point-of-site testing (POST) methods suitable for novice end users that can be validated and internationally accepted remain an objective for both regulators and the industry. The range of commercial testing kits on the market for marine toxin analysis remains limited and even more so those meeting the requirements for use in regulatory control. Individual assays include enzyme-linked immunosorbent assays (ELISA) and lateral flow membrane-based immunoassays (LFIA) for EU-regulated toxins, such as okadaic acid (OA) and dinophysistoxins (DTXs), saxitoxin (STX) and its analogues and domoic acid (DA) in the form of three separate tests offering varying costs and benefits for the industry. It can be observed from the literature that not only are developments and improvements ongoing for these assays, but there are also novel assays being developed using upcoming state-of-the-art biosensor technology. This review focuses on both currently available methods and recent advances in innovative methods for marine biotoxin testing and the end-user practicalities that need to be observed. Furthermore, it highlights trends that are influencing assay developments such as multiplexing capabilities and rapid POST, indicating potential detection methods that will shape the future market.

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Preparation of Monoclonal Antibody for Brevetoxin 1 and Development of Ic-ELISA and Colloidal Gold Strip to Detect Brevetoxin 1

Cited by 22 publications

References 24 publications

Marine Toxins Detection by Biosensors Based on Aptamers

Marine Toxins Detection by Biosensors Based on Aptamers

Biological characterisation and application of human MTH1 and monoclonal antibody preparation

Current Trends and Challenges for Rapid SMART Diagnostics at Point-of-Site Testing for Marine Toxins

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