Kim Shin-Hee, Huang Tung-Shi, Seymour Thomas A, Wei Cheng-I, Kempf Stephen C, Bridgman C Roger, Momcilovic Dragan, Clemens Roger A, An Haejung
Department of Nutritional Sciences, College of Human Environmental Sciences, Oklahoma State University, Stillwater, Oklahoma 74078, USA.
J Food Prot. 2005 Sep;68(9):1860-5. doi: 10.4315/0362-028x-68.9.1860.
An immunoassay system was developed for efficient detection of prohibited meat and bone meal (MBM) in animal feed. Monoclonal antibodies (MAbs) were raised against bovine smooth muscle autoclaved at 130 degrees C for 20 min. Among the 1,500 supernatants of hybridoma cells screened, MAbs 3E1, 1G3, and 3E10 were selected and characterized in this study. The first set of MAbs produced, 3E1 and 1G3, had stronger reactivity against MBM than against smooth muscle that was heat treated at 90 degrees C for 10 min. However, reactivity gradually increased against smooth muscle that was autoclaved at 130 degrees C for up to 1 h. The enzyme-linked immunosorbent assay for detection of MBM in animal feed was optimized with the MAb 3E10 because of its superior performance. MAb 3E10 diluted to 100-fold was used to differentiate bovine MBM from that of other species in ingredients used for commercial animal feeds and could detect down to 0.05% MBM mixed in animal feed.
开发了一种免疫分析系统，用于高效检测动物饲料中违禁的肉骨粉（MBM）。制备了针对在130℃高压灭菌20分钟的牛平滑肌的单克隆抗体（MAb）。在筛选的1500个杂交瘤细胞上清液中，本研究选择并鉴定了单克隆抗体3E1、1G3和3E10。产生的第一组单克隆抗体3E1和1G3对肉骨粉的反应性比对在90℃热处理10分钟的平滑肌更强。然而，对在130℃高压灭菌长达1小时的平滑肌的反应性逐渐增加。由于其卓越的性能，用单克隆抗体3E10优化了用于检测动物饲料中肉骨粉的酶联免疫吸附测定。稀释至100倍的单克隆抗体3E10用于区分商业动物饲料所用成分中牛的肉骨粉与其他物种的肉骨粉，并且能够检测出混入动物饲料中低至0.05%的肉骨粉。
