MJAS

Manuscript Details

Development of Antibody-based Kit for the Detection of Ochratoxin A in Grain Corn

Noor, S.J., Norhafniza, A., Adlin, A.A.K., Faridah, S. and Nur, A.M.S.*

Biotechnology and Nanotechnology Research Centre, Malaysian Agricultural Research and Development Institute (MARDI), Persiaran MARDI-UPM, 43400 Serdang, Selangor, Malaysia

Malaysian J. Anim. Sci. 2021 24(2): 66-77

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The contamination of ochratoxin A (OA) in animal feed has become a huge threat to livestock and husbandry industries globally. Many countries had faced overwhelmed economic loss due to low feed and animal output productivities. This study reported the in-house production and characterisation of antibody IgG against OA. The performance of IgGs collected from two rabbits were investigated under titer and protein assay experimentation. Most IgGs exhibited good titer performance with high protein amount, especially during 2nd and 3rd bleeds. Further study was carried out using antibodies from the O1 3rd bleed. Direct competitive assay of OA antibody in phosphate buffer saline (PBS) and corn matrix system was conducted at different OA standard concentrations. Good linearity (R 2 = 0.9533 in PBS and R 2 = 0.9903 in corn matrix) with a broad working range were obtained from both systems. The sensitivity of the developed optical immunodiagnostic was confirmed with a low limit of detection (LOD) achieved, which was lower than the permitted limit of 5 ppb (LOD = 4.03 ppb in PBS and LOD = 2.70 ppb in corn matrix). Intra and inter-species cross-reaction showed that the produced OA antibody was highly selective towards its specific target analyte. The developed immunodiagnostic was verified with chromatographic analysis and the results from both detection methods complemented each other.