Comparative immunochemical characterization of polyclonal and monoclonal antibodies to aflatoxin B1

Ma, Burkin; Iakovleva,; Vv, Sviridov; Aa, Burkin; Gp, Kononenko; Ha, Soboleva

doi:10.1007/bf02731897

Cited by 5 publications

(5 citation statements)

References 5 publications

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“…The average IC50 value calculated from the standard curves obtained from six consecutive measurements is 90 pg mL −1 , while the limit of detection (LOD) of the ELISA based on three times the standard deviation (3 × SD) at zero concentration is 18 pg mL −1 . Many mAb (or pAb)-based dc-ELISAs or ic-ELISAs for the detection of AFB1 in different samples have been developed in recent decades (Table 1) [20][21][22][23][24][25][26][27][28][29][30][31][32][33].…”

Section: Sensitivity Of Elisamentioning

confidence: 99%

“…For AFB 2 , although there is no an olefinic bond in the furan ring, due to the high similarity in the molecular structure between AFB 1 and AFB 2 , the CR of the mAb with AFB 2 was as much as 23.6%. Many mAb (or pAb)-based dc-ELISAs or ic-ELISAs for the detection of AFB1 in different samples have been developed in recent decades (Table 1) [20][21][22][23][24][25][26][27][28][29][30][31][32][33].…”

Section: Specificity Of the Elisamentioning

confidence: 99%

“…In recent decades, many immunoassays, including chemiluminescence immunoassay [16], time-resolved fluorescence immunoassay [17], electrochemical immunosensors [18], and immunochromatographic assay [19], have been reported for the detection of AFB 1 . Furthermore, many direct competitive (dc)-or indirect competitive (ic)-ELISAs based on polyclonal antibodies (pAbs) or monoclonal antibodies (mAbs) for detecting AFB 1 have been developed (Table 1) [20][21][22][23][24][25][26][27][28][29][30][31][32][33]. However, establishing a sensitive immunoassay is mainly dependent upon the quality of antibody.…”

Section: Introductionmentioning

confidence: 99%

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A Highly Sensitive and Group-Specific Enzyme-Linked Immunosorbent Assay (ELISA) for the Detection of AFB1 in Agriculture and Aquiculture Products

Cao,

Wang,

et al. 2024

Molecules

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Aflatoxins (AFs) including AFB1, AFB2, AFG1 and AFG2 are widely found in agriculture products, and AFB1 is considered one of the most toxic and harmful mycotoxins. Herein, a highly sensitive (at the pg mL−1 level) and group-specific enzyme-linked immunosorbent assay (ELISA) for the detection of AFB1 in agricultural and aquiculture products was developed. The AFB1 derivative containing a carboxylic group was synthesized and covalently linked to bovine serum albumin (BSA). The AFB1-BSA conjugate was used as an immunogen to immunize mice. A high-quality monoclonal antibody (mAb) against AFB1 was produced by hybridoma technology, and the mAb-based ELISA for AFB1 was established. IC50 and limit of detection (LOD) of the ELISA for AFB1 were 90 pg mL−1 and 18 pg mL−1, respectively. The cross-reactivities (CRs) of the assay with AFB2, AFG1, and AFG2 were 23.6%, 42.5%, and 1.9%, respectively, revealing some degree of group specificity. Corn flour, wheat flour, and crab roe samples spiked with different contents of AFB1 were subjected to ELISA procedures. The recoveries and relative standard deviation (RSD) of the ELISA for AFB1 in spiked samples were 78.3–116.6% and 1.49–13.21% (n = 3), respectively. Wheat flour samples spiked with the mixed AF (AFB1, AFB2, AFG1, AFG2) standard solution were measured by ELISA and LC-MS/MS simultaneously. It was demonstrated that the proposed ELISA can be used as a screening method for evaluation of AFs (AFB1, AFB2, AFG1, AFG2) in wheat flour samples.

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Section: Sensitivity Of Elisamentioning

confidence: 99%

Section: Specificity Of the Elisamentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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A Highly Sensitive and Group-Specific Enzyme-Linked Immunosorbent Assay (ELISA) for the Detection of AFB1 in Agriculture and Aquiculture Products

Cao,

Wang,

et al. 2024

Molecules

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“…ELISA has advantages for rapid screening of aflatoxins from many kinds of matrix, and the detection limits of ELISA assay can be comparable with instrumental assay. The usefulness of these immunoassays is dependent on the specificity and sensitivity of antibody used, so the production of mAb and polyclonal antibodies to AFB1 and their application in immunoassay have been reported (Burkin et al, 2000 ; Mortimer et al , 1988 ; Saha et al , 2007) . Here, we produced a new anti-AFB1 mAb with high affinity to natural aflatoxin B1 and its analogs, then applied it to a direct competitive antibody coated ELISA (Ab-DC-ELISA) and a direct competitive antigen coated ELISA (Ag-DC-ELISA).…”

Section: Introductionmentioning

confidence: 99%

Production of Group Specific Monoclonal Antibody to Aflatoxins and its Application to Enzyme-linked Immunosorbent Assay

Kim¹,

Cha²,

Bischoff³

et al. 2011

Toxicological Research

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Through the present study, we produced a monoclonal antibody against aflatoxin B1 (AFB1) using AFB1- carboxymethoxylamine BSA conjugates. One clone showing high binding ability was selected and it was applied to develop a direct competitive ELISA system. The epitope densities of AFB1-CMO against BSA and KLH were about 1 : 6 and 1 : 545, respectively. The monoclonal antibody (mAb) from cloned hybridoma cell was the IgG1 subclass with λ-type light chains. The IC50s of the monoclonal antibody developed for AFB1, AFB2, AFG1 and AFG2 were 4.36, 7.22, 6.61 and 29.41 ng/ml, respectively, based on the AFB1-KLH coated ELISA system and 15.28, 26.62, 32.75 and 56.67 ng/ml, respectively, based on the mAb coated ELISA. Cross-relativities of mAb to AFB1 for AFB2, AFG1 and AFG2 were 60.47, 65.97 and 14.83% in the AFB1-KLH coated ELISA, and 59.41, 46.66 and 26.97% in the mAb coated ELISA, respectively. Quantitative calculations for AFB1 from the AFB1-Ab ELISA and AFB1-Ag ELISA ranged from 0.25 to 25 ng/ml (R2 > 0.99) and from 1 to 100 ng/ml (R2 > 0.99), respectively. The intra- and inter-assay precision CVs were < 10% in both ELISA assay, representing good reproducibility of developed assay. Recoveries ranged from 79.18 to 91.27%, CVs ranged from 3.21 to 7.97% after spiking AFB1 at concentrations ranging from 5 to 50 ng/ml and following by extraction with 70% methanol solution in the Ab-coated ELISA. In conclusion, we produced a group specific mAb against aflatoxins and developed two direct competitive ELISAs for the detection of AFB1 in feeds based on a monoclonal antibody developed.

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“…E-mail: kongqing@ouc.edu.cn Tel: +86-532-8203-1851. Fax: +86-532-8203-2389 Abbreviation: AFs, Aflatoxins. especially dangerous to human and animal health because of their high hepatotoxicity and nephrotoxicity, as well as their carcinogenic and genotoxic effects (Burkin et al, 2000).…”

Section: Introductionmentioning

confidence: 99%

Statistical experimental methods for optimizing the cultivating conditions for Rhodococcus erythropolis

Zhai¹,

Kong²,

Guan³

et al. 2011

Afr. J. Biotechnol.

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Rhodococcus erythropolis was found to effectively degrade aflatoxin B l produced by Aspergillus flavus and Aspergillus parasiticus. However, one problem of concern was the slow growth of this strain. In this study, Plackett-Burman design was used to select the most important variables, namely, temperature, pH, inoculum size, liquid volume, agitation speed and culture time that affected the growth of R. erythropolis. Central composite experimental design and response surface analysis were adopted to derive a statistical model for optimizing the culture conditions. From the obtained results, it can be concluded that the optimum parameters were: temperature, 15.3°C; pH, 5.56; inoculum size, 4%; liquid volume, 70 ml in 250 ml flask; agitation speed, 180 rpm; and culture time, 58.2 h. At this optimum point, the populations of the viable organisms could reach 10 8 colony forming units (CFU)/ml, which was 100 times higher than that incubated under the initial conditions. After 58.2 h incubation in this optimum cultivating conditions, 53.9 ± 2.1% of aflatoxin B 1 was degraded, while only 20.6±1.4% of aflatoxin B 1 was degraded in the initial conditions.

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Comparative immunochemical characterization of polyclonal and monoclonal antibodies to aflatoxin B1

Cited by 5 publications

References 5 publications

A Highly Sensitive and Group-Specific Enzyme-Linked Immunosorbent Assay (ELISA) for the Detection of AFB1 in Agriculture and Aquiculture Products

A Highly Sensitive and Group-Specific Enzyme-Linked Immunosorbent Assay (ELISA) for the Detection of AFB1 in Agriculture and Aquiculture Products

Production of Group Specific Monoclonal Antibody to Aflatoxins and its Application to Enzyme-linked Immunosorbent Assay

Statistical experimental methods for optimizing the cultivating conditions for Rhodococcus erythropolis

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