J Vet Sci.  2012 Dec;13(4):363-369.

A novel mycotoxin purification system using magnetic nanoparticles for the recovery of aflatoxin B1 and zearalenone from feed

• Kim HJ
• Kim SH
• Lee JK
• Choi CU
• Lee HS
• Kang HG
• Cha SH

Abstract

In this study, we developed a novel tool for purifying two mycotoxins, aflatoxin B1 (AFB1) and zearalenone (ZEN), in feed. This system utilized monoclonal antibodies (mAbs) against AFB1 and ZEN, and magnetic nanoparticles (MNPs). Among ten MNPs with different diameters and functional groups, a 100-nm diameter MNP (fMA) conjugated to an amine group (-NH2) was found to be optimum for coupling with mAbs. The optimal mAb concentrations for coupling to the fMA along with mycotoxin purification capacities of the fMA-mAb conjugates (fMA-AFB1 and fMA-ZEN) were determined. A comparison of mean recovery rates (from corn and product X feed) between the fMA-mAb conjugates and immunoaffinity columns (IAC-AFB1 and IAC-ZEN) showed that the rate for fMA-AFB1 (90~92% and 81~88%) was higher (p > 0.05) than that of IAC-AFB1 (81~84% and 72~78%) for AFB1 (5, 10, 15 ng/mL), and the rate for fMA-ZEN (99~100% and 92~94%) was significantly higher (p < 0.01) than that of IAC-ZEN (86~88% and 81~88%) for ZEN (10, 25, 50 ng/mL) except at a concentration of 10 ng/mL, demonstrating the remarkable purification efficiency of the novel fMA-mAb method. Additionally, mycotoxin purification was much faster using our novel method (approx. 5 min) than the IAC-based technique (> 30 min). This study suggests that the novel purification system we developed would be a useful tool for monitoring and regulating mycotoxin contamination in feed, and replace IAC methods.

Keyword

aflatoxin B1; immunoaffinity column; mAb; magnetic nanoparticle; zearalenone

MeSH Terms

Aflatoxin B1
Antibodies, Monoclonal
Magnetics
Magnets
Mycotoxins
Nanoparticles
Zea mays
Zearalenone

Figure

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  Fig. 1 Competitive ELISA for measuring mAb specificity and affinity of (A) kj-AFB and (B) kk-ZEN. The mAbs we developed (kj-AFB or kk-ZEN) or commercial mAbs [C] at different dilutions (1/250, 1/500, and 1/1,000 for C; 1/64, 1/128, 1/256, 1/512, and 1/1024 for kk-ZEN; 1/16, 1/32, 1/64, 1/128, and 1/256 for kj-AFB). kj-AFB and kk-ZEN were found to have greater specificity as well as affinity than the commercial mAbs.

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  Fig. 2 Separation of MNPs using a magnet.

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  Fig. 3 Coupling efficiency of MNPs with a diameter of 100 nm. MNPs (2 mg) were coupled with 100 µg of the kk-ZEN mAb specific for ZEN (n = 3). The amount of bound mAb is presented as the mean ± SE. *p < 0.01.

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  Fig. 4 Optimal coupling conditions with different concentration of mAbs and the mycotoxin purification capacity. MNPs (2 mg) were coupled with 50, 100, or 200 µg of mAbs (n = 3) and 1 mg each fluidMAG-amine (fMA)-AFB1 (A) and fMA-ZEN (B). All values were significantly different (p < 0.01) except for differences in the amounts of mycotoxin purified by MNPs coupled with 50 µg or 100 µg of AFB1-specific mAb. *Indicates the mean ± SE.

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  Fig. 5 Comparison of recovery rates for the MNP-mAb conjugates (fMA-AFB1 and fMA-ZEN) and immunoaffinity columns (IAC-AFB1 and IAC-ZEN) from corn (A) or product X (B) feed samples (n = 3). Recovery rate (%) = (amount of purified mycotoxins/amount of spiked mycotoxins) × 100. *p < 0.01.

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