燕窝唾液酸糖蛋白间接竞争酶联免疫检测方法的建立  

Establishment of indirect competitive enzyme linked immunosorbent assay for detection of sialoglycoprotein of edible bird’s nest

作　　者：何金麟 王仍瑞 张世伟[2] 冯荣虎[2] 范群艳 柳训才 徐敦明 HE Jin-Lin;WANG Reng-Rui;ZHANG Shi-Wei;FENG Rong-Hu;FAN Qun-Yan;LIU Xun-Cai;XU Dun-Ming(Technical Center of Xiamen Customs,Xiamen 361026,China;Shenzhen Academy of Metrology and Quality Inspection,Shenzhen 518102,China;Yan Palace Seelong Food Co.,Ltd.,Xiamen 361100,China)

机构地区：[1]厦门海关技术中心,厦门361026 [2]深圳市计量质量检测研究院,深圳518102 [3]厦门市燕之屋丝浓食品有限公司,厦门361100

出　　处：《食品安全质量检测学报》2022年第23期7630-7636,共7页Journal of Food Safety and Quality

基　　金：海关总署科研项目(2021HK198)。

摘　　要：目的建立间接竞争酶联免疫(indirect competitive enzyme linked immunesorbent assay,IC-ELISA)检测燕窝中唾液酸糖蛋白的分析方法。方法以唾液酸糖蛋白为抗原,制备单克隆抗体,对IC-ELISA步骤中相关参数进行优化,确定最佳反应条件。结果最佳包被抗原为1:2000,抗体稀释倍数为1:5000;封闭液浓度为1%牛血清白蛋白(bovine serum albumin,BSA),封闭时间为150 min;竞争时间30 min;酶标二抗浓度稀释倍数为1:1000,孵育时间30 min;显色时间为10 min。建立的IC-ELISA检测方法半抑制浓度(50%inhibition concentration,IC_(50))为8.61μg/mL,线性范围为2.23~33.25μg/mL,回收率为82.7%~92.0%,相对标准偏差(relative standard deviation,RSDs)为1.7%~8.9%。结论该方法灵敏度较高、特异性强,可以有效地鉴别假冒伪劣燕窝产品,具有较好的实际应用价值。Objective To establish an indirect competitive enzyme linked immunosorbent assay(IC-ELISA)method for the detection of sialoglycoprotein in edible bird’s nest.Methods Monoclonal antibody was prepared with sialoglycoprotein as antigen,and the relevant parameters in IC-ELISA step were optimized to determine the optimal reaction conditions.Results The optimal coating antigen was determined to be 1:2000,and the dilution of antibody was 1:5000.The concentration of blocking liquid was 1%bovine serum albumin(BSA)and the blocking time was 150 min.Competition time was 30 min.The concentration dilution of enzyme-conjugate secondary antibody was 1:1000.The incubation time was 30 min and colouration time was 10 min.The results showed that the 50%inhibition concentration(IC_(50))of the established IC-ELISA was 8.61μg/mL.The linear range was 2.23‒33.25μg/mL and the recoveries were between 82.7%‒92.0%,with relative standard deviations(RSDs)of 1.7%–8.9%.Conclusion This method has high sensitivity and specificity,which can be used for effectively identify fake bird’s nest products and has good practical application value.

关 键 词：燕窝 唾液酸糖蛋白 间接竞争酶联免疫分析法 

分 类 号：TS218[轻工技术与工程—粮食、油脂及植物蛋白工程]