薛海燕,吴剑,张颖,宋宏新,贺宝元.牛β-乳球蛋白胶体金免疫层析试纸条的研制及检测应用[J].,2021,37(3):259-266.
牛β-乳球蛋白胶体金免疫层析试纸条的研制及检测应用
Development and Application of Bovine β-lactoglobulin Gold Immunoassay Strip
投稿时间:2020-09-14  
DOI:10.13982/j.mfst.1673-9078.2021.3.0862
中文关键词:  牛β-乳球蛋白  β-lg单克隆抗体  免疫层析  市售羊乳粉  掺伪
英文关键词:bovine β-lactoglobulin  β-lg monoclonal antibody  immunochromatography  commercially available sheep milk powder  adulteration
作者简介:薛海燕(1979-),女,博士,副教授,研究方向:食品加工与质量控制技术
基金项目:陕西省重点研发计划项目(2020ZDLNY02-09);陕西省教育厅服务地方专项项目(19JC05);西安市科技计划农业科技创新工程项目(20193035YF023NS023;20NYYF0016)
作者单位
薛海燕 (1.陕西科技大学食品与生物工程学院,陕西西安 710021) 
吴剑 (1.陕西科技大学食品与生物工程学院,陕西西安 710021) 
张颖 (1.陕西科技大学食品与生物工程学院,陕西西安 710021) 
宋宏新 (1.陕西科技大学食品与生物工程学院,陕西西安 710021) 
贺宝元 (2.陕西科技大学轻工科学与工程学院,陕西西安 710021) 
AuthorInstitution
XUE Hai-yan (1.Institute of Food Biotechnology, Shaanxi University of Science and Technology of Science and Technology, Xi’an 710021, China) 
WU Jian (1.Institute of Food Biotechnology, Shaanxi University of Science and Technology of Science and Technology, Xi’an 710021, China) 
ZHANG Ying (1.Institute of Food Biotechnology, Shaanxi University of Science and Technology of Science and Technology, Xi’an 710021, China) 
SONG Hong-xin (1.Institute of Food Biotechnology, Shaanxi University of Science and Technology of Science and Technology, Xi’an 710021, China) 
HE Bao-yuan (2.Institute of Light Industry Science and Engineering, Shaanxi University of Science and Technology of Science and Technology, Xi’an 710021, China) 
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中文摘要:
      研制了一种可快速检测羊乳及制品中牛源β-乳球蛋白(β-lactoglobulin,β-lg)的胶体金免疫层析试纸条。通过杂交瘤技术制备牛β-lg特异性单克隆抗体(monoclonal antibody,mAb),利用柠檬酸钠还原氯金酸,形成30 nm胶体金颗粒,并用于标记牛β-lgmAb。采用竞争法研制免疫层析试纸条,将胶体金标记的牛β-lgmAb包被于金标垫,牛β-lg和羊抗鼠IgG标记于硝酸纤维素膜分别作为检测线(T线)和质控线(C线),牛β-lg和二抗的最佳包被浓度均为1.0 mg/mL。制得的单克隆抗体纯度都在90%以上,效价均在10000以上且特异性较好。该试纸条对牛β-lg的检测限(limit of detection,LOD)值为3.13 μg/mL,对牛源α-CN,牛源β-CN,牛源κ-CN,牛源α-LA,BSA均未产生交叉反应,对脱脂羊乳粉中掺杂脱脂牛奶粉的LOD值为5%,并用该方法对市售羊奶及配方羊奶粉进行分析,检测结果与商品化的ELISA试剂盒一致。该方法前处理快速简单,可在5 min内对牛β-lg进行检测,可用于羊乳制品的现场快速检测。
英文摘要:
      A colloidal gold immunochromatography method for the rapid determination of β-lactoglobulin (β-lg) in goat milk and its products were established. Bovine β-lg specific monoclonal antibody (monoclonal antibody, mAb) was prepared by hybridoma technique. Sodium citrate was used to reduce chloroauric acid. The 30 nm colloidal gold particles were formed and used to label bovine β-lgmAb. The competition method was used to develop immunochromatographic strips. A bovine β-lgmAb bag labeled with colloidal gold was placed on the gold label pad. Bovine β-lg and sheep anti-mouse IgG were labeled on nitrocellulose film as detection line (T line) and quality control line (C line), respectively. The optimal encapsulation concentration of bovine β-lg and secondary antibodies were both 1.0 mg/mL. The purity and titer of monoclonal antibody were all above 90% and above 10000 with high specificity. The limit of detection (LOD) of bovine β-lg was 3.13 μg/mL. There was no cross-reaction against other substrate components (bovine α-CN, β-CN, κ-CN, α-LA, BSA). The LOD of whole fat goat milk powder adulterated with skimmed cow milk powder was 0.5 %. The method was used to analyze the commercial goat milk and formula goat milk powder. The results of this method were consistent with that of the commercial ELISA kit. This method could be used to detect bovine β-lg in 5 min, and could be used to on-site rapid detection of sheep milk products.
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