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Immunoassay for the Detection of Animal Central Nervous Tissue in Processed Meat and Feed Products

Rao, Qinchun, Richt, Juergen A., Hsieh, Yun-Hwa Peggy
Journal of agricultural and food chemistry 2016 v.64 no.18 pp. 3661-3668
Western blotting, autoclaving, beef, brain, cattle, cross reaction, detection limit, enzyme-linked immunosorbent assay, feeds, hybridomas, monoclonal antibodies, myelin basic protein, pork, processed foods, processed meat, soybean meal
An indirect competitive enzyme-linked immunosorbent assay (icELISA) based on the detection of the thermal-stable central nervous tissue (CNT) marker protein, myelin basic protein (MBP), was developed to detect animal CNT in processed meat and feedstuffs. Two meat samples (cooked at 100 °C for 30 min and autoclaved at 133 °C for 20 min) of bovine brain in beef and two feed samples (bovine brain meal in beef meal and in soybean meal) were prepared at levels of 0.0008, 0.0031, 0.0063, 0.0125, 0.025, 0.05, 0.1, 0.2, 0.4, 0.8, and 1.6%. An anti-MBP monoclonal antibody (mAb3E3) was produced using the hybridoma technique and characterized using Western blot. The optimized icELISA was CNT-specific without cross-reactivity with either meat (beef and pork) or soybean meal samples and had low intra-assay (%CV ≤ 3.5) and interassay variability (%CV ≤ 3.3), with low detection limits for bovine MBP (6.4 ppb) and bovine CNT spiked in both meat (0.05%) and feed (0.0125%) samples. This assay is therefore suitable for the quantitative detection of trace amounts of contaminated animal CNT in processed food and feed products.