Authors: S-H Cha, J-K Lee, H-S Jung, S-H Kim, H-J Kim, C-U Choi, H-G Kang
Affilation: National Veternary Research and Quarantine Service, Korea
Pages: 569 - 571
Keywords: magnetic nanoparticles
Introduction: A common method for determination of mycotoxins contaminated in feeds combines immunoaffinity chromatography (IAC) and high-performance liquid chromatography (HPLC) with UV detection1,2. However, the IAC method to collect mycotoxins is expensive and time-consuming. Moreover, its collection efficiency has failed to provide complete satisfaction for users. The present study was conducted to develop the better collection system utilizing magnetic nanoparticle (MNP), which is synthesized for the present study and featured with easy collection by magnetism and fast dispersibility by tiny size. Materials and Methods: MNP was synthesized by a slight modification of the literature method,3 and the surface of MNP was coated with silica (SiO2) by a polyvinylpyrrolidone(PVP)-mediated method to generate MNP@SiO2 as previously reported in the literature.4 Finally, the surface of MNP@SiO2 was further modified with aminoorganosilanes in order to introduce amino-terminal groups for simple and easy coupling with biomolecules. Monoclonal antibodies, which were kj-AFB and kk-ZEN against aflatoxin B1(AFB1) and zearalenone (ZEN), respectively, were produced in our laboratory, and covalently coupled with the synthesized MNP. Selective reaction and harvest were carried out easily by re-suspension and magnetic separation. The recovery efficiency for mycotoxin in feed was compared between AFB1 or ZEN mAb coupled MNP and commercial AFB1 or ZEN-IACs. Results and Discussion: The optimal amount of covalently coupled mAb to MNP (C-AFB1 mAb/MNP and C-ZEN mAb/MNP) was 42.16ug and 40.39ug to 1mg of MNP respectively. C-AFB1 mAb/MNP and C-ZEN mAb/MNP showed individual binding capacity of 1.03ng and 2.24ng per 1ug mAb, respectively. In application for collection of mycotoxins (10ppb for AFB1 and 50ppb for ZEN) spiked into feed, 45ug C-AFB1 mAb/MNP and 40.0ug C-ZEN mAb/MNP were used to recover all mycotoxins. Comparison of recovery efficiency showed that C-AFB1/MNP and AFB1-IAC were 90.28% and 83.7%, respectively, while C-ZEN/MNP and ZEN-IAC were 99.9% and 86.0% respectively, demonstrating remarkable efficiency of the collection system using nanoparticles for mycotoxin from feed. Conclusion: It is suggested that the novel collection system would be a great tool to monitor and regulate mycotoxin contaminated in feeds. References 1. Visconti A. et al.1998. Journal of Chromatogaphy A. Jul 31; 815(1):133-40. 2. Castegnaro M. et al. 2006. Molecular Nutrition and Food Research. May; 50(6):480-7. 3. Deng H. et al. 2005. Angewandte Chemie International Edition. Apr 30; 44(18): 2782-2785. 4. Yoon T.-J. et al. 2005. Angewandte Chemie International Edition. Feb 30; 44(7): 1068-1071.