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

• Published in: Journal of veterinary science (Suwŏn-si, Korea) Vol. 16; no. 4; pp. 431 - 437
• Main Authors: Kim, Nam-Gun, Kim, Myeong-Ae, Park, Young-Il, Jung, Tae-Sung, Son, Seong-Wan, So, ByungJae, Kang, Hwan-Goo
• Format: Journal Article
• Language: English
• Published: Korea (South) 대한수의학회 2015; The Korean Society of Veterinary Science
• Subjects: Animals; Antibodies, Monoclonal - chemistry; Antineoplastic Agents - analysis; Eggs - analysis; Enzyme-Linked Immunosorbent Assay - methods; Female; Fluoroquinolones - analysis; Food Contamination - analysis; Magnetite Nanoparticles - chemistry; Meat - analysis; Mice; Mice, Inbred BALB C; Original; 수의학; magnetic nanoparticle; enrofloxacin; monoclonal antibody; enzyme-linked immunosorbent assay
• ISSN: 1229-845X; 1976-555X
• DOI: 10.4142/jvs.2015.16.4.431

Monoclonal anti-enrofloxacin antibody was prepared for a direct competitive enzyme-linked immunosorbent assay (ELISA) and purification system using monoclonal antibody (mAb) coupled magnetic nanoparticles (MNPs). The IC50 values of the developed mAb for enrofloxacin (ENR), ciprofloxacin, difloxacin, sarafloxacin, pefloxacin, and norfloxacin were 5.0, 8.3, 9.7, 21.7, 36.0, and 63.7 ng/mL, respectively. The lowest detectable level of ENR was 0.7 ng/mL in the prepared ELISA system. To validate the developed ELISA in the food matrix, known amounts of ENR were spiked in meat and egg samples at 10, 20 and 30 ng/mL. Recoveries for ENR ranged from 72.9 to 113.16% with a coefficient of variation (CV) of 2.42 to 10.11%. The applicability of the mAb-MNP system was verified by testing the recoveries for ENR residue in three different matrices. Recoveries for ENR ranged from 75.16 to 86.36%, while the CV ranged from 5.08 to 11.53%. Overall, ENR-specific monoclonal antibody was prepared and developed for use in competitive to ELISAs for the detection of ENR in animal meat samples. Furthermore, we suggest that a purification system for ENR using mAb-coupled MNPs could be useful for determination of ENR residue in food.