Broad-specificity ELISA with a heterogeneous strategy for sensitive detection of microcystins and nodularin

Lü, Ning; Ling, Li; Guan, Tian; Wang, Lanteng; Wang, Dian; Zhou, Jian; Ruan, Ting; Shen, Xing; Li, Xiangmei; Sun, Yuanming; Lei, Hongtao

doi:10.1016/j.toxicon.2019.12.003

Cited by 26 publications

(11 citation statements)

References 38 publications

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“…New ELISA utilizing antibodies extracted from eggs of immunized chickens, competitive indirect ELISA (CI-ELISA) utilizing antibodies raised in sheep against 6(E) Adda, as well as competitive direct ELISA (CD-ELISA) and CI-ELISA generated from rabbits conjugated with gamma-globulin were applied to detected various MC variants in water samples [ 43 , 48 , 126 ]. Recently, the newly developed CI-ELISA, which utilized MC-LR-keyhole limpet hemocyanin (KLH) for New Zealand white rabbit immunization and produced antibodies, detected MC-LR with a limit of detection (LOD) of 0.0016 ng/mL [ 107 ].…”

Section: Analytical Methods To Detect Microcystinsmentioning

confidence: 99%

“…A small amount of water sample is needed for toxins identification. Generally, ELISA is capable of yielding repeatability, reproducibility and variability results of MCs concentrations compared to the other methods [ 86 , 87 , 107 , 118 ]. Besides, no sample cleanup is needed, detection limits are often below the WHO’s 1 µg/L guideline value, and it is sensitive to low pH (formic acid), MeOH or MeCN [ 79 , 87 , 88 , 114 , 115 ].…”

Section: Analytical Methods To Detect Microcystinsmentioning

confidence: 99%

“…Interestingly an ELISA antibody developed against specific MC variant(s) may give false negatives if used to detect other MC variant(s) to which it is not sensitive to. The exact MC content and toxicity are likely to not be detected due to the variations in the specificity of the antibody [ 43 , 48 , 107 , 126 ]. Even though ELISA is capable of detecting MCs in solid samples (including fish tissues), the extraction methods it uses (for the solid samples) are not typically suitable for field application (non-solid samples) [ 82 , 115 , 119 ].…”

Section: Analytical Methods To Detect Microcystinsmentioning

confidence: 99%

See 2 more Smart Citations

A Mini-Review on Detection Methods of Microcystins

Massey

Jia

et al. 2020

Toxins

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Cyanobacterial harmful algal blooms (CyanoHABs) produce microcystins (MCs) which are associated with animal and human hepatotoxicity. Over 270 variants of MC exist. MCs have been continually studied due of their toxic consequences. Monitoring water quality to assess the presence of MCs is of utmost importance although it is often difficult because CyanoHABs may generate multiple MC variants, and their low concentration in water. To effectively manage and control these toxins and prevent their health risks, sensitive, fast, and reliable methods capable of detecting MCs are required. This paper aims to review the three main analytical methods used to detect MCs ranging from biological (mouse bioassay), biochemical (protein phosphatase inhibition assay and enzyme linked immunosorbent assay), and chemical (high performance liquid chromatography, liquid chromatography-mass spectrometry, high performance capillary electrophoresis, and gas chromatography), as well as the newly emerging biosensor methods. In addition, the current state of these methods regarding their novel development and usage, as well as merits and limitations are presented. Finally, this paper also provides recommendations and future research directions towards method application and improvement.

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Section: Analytical Methods To Detect Microcystinsmentioning

confidence: 99%

Section: Analytical Methods To Detect Microcystinsmentioning

confidence: 99%

Section: Analytical Methods To Detect Microcystinsmentioning

confidence: 99%

See 1 more Smart Citation

A Mini-Review on Detection Methods of Microcystins

Massey

Jia

et al. 2020

Toxins

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“…They are often aggravated by the low immunogenicity or non-uniqueness of the selected structures and, moreover, require complex preliminary chemical synthetic work. An alternative approach to competitive immunoassays is to use different antigen derivatives in immunization and analysis [19][20][21][22][23][24][25][26][27][28]. This so-called "heterologous" immunoassay ensures that not all antibodies produced are involved in the competitive analytical interactions, and the spectrum of selectivity is thereby narrowed in comparison with the traditional "homologous" assay.…”

Section: Introductionmentioning

confidence: 99%

Changing Cross-Reactivity for Different Immunoassays Using the Same Antibodies: Theoretical Description and Experimental Confirmation

et al. 2021

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Many applications of immunoassays involve the possible presence of structurally similar compounds that bind with antibodies, but with different affinities. In this regard, an important characteristic of an immunoassay is its cross-reactivity: the possibility of detecting various compounds in comparison with a certain standard. Based on cross-reactivity, analytical systems are assessed as either high-selective (responding strictly to a specific compound) or low-selective (responding to a number of similar compounds). The present study demonstrates that cross-reactivity is not an intrinsic characteristic of antibodies but can vary for different formats of competitive immunoassays using the same antibodies. Assays with sensitive detection of markers and, accordingly, implementation at low concentrations of antibodies and modified (competing) antigens are characterized by lower cross-reactivities and are, thus, more specific than assays requiring high concentrations of markers and interacting reagents. This effect was confirmed by both mathematical modeling and experimental comparison of an enzyme immunoassay and a fluorescence polarization immunoassay of sulfonamides and fluoroquinolones. Thus, shifting to lower concentrations of reagents decreases cross-reactivities by up to five-fold. Moreover, the cross-reactivities are changed even in the same assay format by varying the ratio of immunoreactants’ concentrations and shifting from the kinetic or equilibrium mode of the antigen-antibody reaction. The described patterns demonstrate the possibility of modulating immunodetection selectivity without searching for new binding reactants.

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“…Actually, except for a few examples, most of the antibodies reported above are broad specificity antibodies originally prepared for MC-LR, but they can recognize also othe MCs as well as NOD-R simultaneously. Previously, with MC-LR-KLH as a common im munogen, multi-residue-recognizing PAbs [16] and MAbs [data not shown] were gener ated in our lab with IC50 of 0.29 µg/L and 18 µg/L for NOD-R, respectively.…”

Section: Introductionmentioning

confidence: 99%

Preparation of Specific Nanobodies and Their Application in the Rapid Detection of Nodularin-R in Water Samples

Yang

et al. 2021

Foods

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Nanobodies have several advantages, including great stability, sensibility, and ease of production; therefore, they have become important tools in immunoassays for chemical contaminants. In this manuscript, nanobodies for the detection of the toxin Nodularin-r (NOD-R), a secondary metabolite of cyanobacteria that could cause a safety risk for drinks and food for its strong hepatotoxicity, were for the first time selected from an immunized Bactrian camel VHH phage display library. Then, a sensitive indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) for NOD-R, based on the nanobody N56 with great thermostability and organic solvent tolerance, was established under optimized conditions. The results showed that the limit of detection for NOD-R was 0.67 µg/L, and the average spike recovery rate was between 84.0 and 118.3%. Moreover, the ic-ELISA method was validated with spiked water sample and confirmed by UPLC–MS/MS, which indicated that the ic-ELISA established in this work is a reproducible detection assay for nodularin residues in water samples.

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Broad-specificity ELISA with a heterogeneous strategy for sensitive detection of microcystins and nodularin

Cited by 26 publications

References 38 publications

A Mini-Review on Detection Methods of Microcystins

A Mini-Review on Detection Methods of Microcystins

Changing Cross-Reactivity for Different Immunoassays Using the Same Antibodies: Theoretical Description and Experimental Confirmation

Preparation of Specific Nanobodies and Their Application in the Rapid Detection of Nodularin-R in Water Samples

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