Magnetic nanoparticle based purification and enzyme-linked immunosorbent assay using monoclonal antibody against enrofloxacin

Kim, Nam-Gun; Kim, Myeong-Ae; Park, Young-Il; Jung, Tae-Sung; Son, Seong-Wan; So, ByungJae; Kang, Hwan-Goo

Conventional methods such as liquid chromatography coupled to various detectors including ultra-violet (UV), mass spectrometry, or fluorescence detection are used for detection of drug residues [2, 31] . These techniques have been shown to be highly specific and sensitive, but such traditional methods require expensive equipment and interpretation of complicated chromatograms or spectral results [13] . Therefore, a rapid, reliable, and easy screening method is required for monitoring of large samples [4] . Enzyme-linked immunosorbent assay (ELISA), which is based on specific antigen-antibody interactions, is the most suitable method for rapid screening of ENR residue in the veterinary field [29, 30] . Monoclonal or polyclonal antibodies have been developed for use in immunochemical detection assays [9, 20] . Many organic solvents or immunoaffinity columns are required to separate FQs from the matrix to enable their analysis. The magnetic nanoparticle (MNP) has emerged for various applications such as gene and drug delivery, treatment of disease, and diagnosis [11, 24] . MNP can bind to different functional groups such as oligonucleotide probes, antibodies, and proteins to produce nanoprobes [19] . Previous studies have indicated the usefulness of nanoparticles for identification of pathogenic bacteria in DNA-microarrays, isolating target organisms from food matrices and screening metal ions in water [8, 17, 26] . Additionally, we reported a rapid purification method using monoclonal antibodies against mycotoxin and MNPs [18] . This study was conducted to develop a direct competitive ELISA system to screen for ENR in foodstuffs and to develop a purification tool for isolating ENR by utilizing the ENR monoclonal antibody (mAb) and MNPs.