食糖中右旋糖酐胶乳增强免疫比浊分析方法的建立  

Establishment of Latex-Enhanced Turbidimetric Immunoassay for Determination of Dextran in Sugar

作　　者：常国炜 刘桂云 罗秀玉 黎志德 农健 黄曾慰 梁达奉 CHANG Guowei;LIU Guiyun;LUO Xiuyu;LI Zhide;NONG Jian;HUANG Zengwei;LIANG Dafeng(Guangdong Engineering Technology Research Center of Enzyme and Biocatalysis,Institute of Biological and Medical Engineering,Guangdong Academy of Sciences,Guangzhou 510316,China;Guangxi Sugar Industry Group Changling Sugar Co.,Ltd.,Fangchenggang 535514,China)

机构地区：[1]广东省科学院生物与医学工程研究所,广东省酶制剂与生物催化工程技术研究中心,广东广州510316 [2]广西糖业集团昌菱制糖有限公司,广西防城港535514

出　　处：《食品科技》2024年第12期288-292,共5页Food Science and Technology

基　　金：广西重点研发计划项目(桂科AB21220054);广东省科学院发展专项资金项目(2022GDASZH-2022010110)。

摘　　要：文章建立了食糖中右旋糖酐胶乳增强免疫比浊分析方法。利用杂交瘤细胞株,通过体内诱生法,制备一定量的右旋糖酐单克隆抗体。通过成本较低的羟基磷灰石层析法,得到纯度为71%的右旋糖酐单克隆抗体。以带有羧基功能基团的胶乳微球作为载体,通过1-乙基-(3-二甲基氨基丙基)碳酰二亚胺介导的化学反应与右旋糖酐单克隆抗体形成酰胺键,实现共价偶联,制备得到右旋糖酐免疫胶乳微球。该方法的反应体系主要为pH7.0的4-羟乙基哌嗪乙磺酸缓冲液,检测仪器为全自动生化分析仪。结果表明,该方法线性关系良好,相关系数为0.9997,检出限为3.68 mg/L。样品的加标回收率为93.8%~105.8%,相对标准偏差为0.50%~1.81%。A latex-enhanced turbidimetric immunoassay for the determination of dextran in sugar was established.A certain amount of dextran monoclonal antibody was prepared using hybridoma cell lines and in vivo induction methods.The dextran monoclonal antibody with a purity of 71%was obtained by low-cost hydroxyapatite chromatography.Carboxylated latex microspheres were used as carriers,and amide bonds were formed between the latex microspheres and dextran monoclonal antibody through a 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide(EDC)-mediated chemical reaction to achieve covalent coupling,resulting in the preparation of dextran immune latex microspheres.The reaction system of this method mainly consists of 2-[4-(2-hydroxyethyl)piperazin-1-yl]ethanesulfonic acid(HEPES)buffer at pH 7.0,and the detection instrument is an automatic biochemical analyzer.The results showed that the method has a good linear relationship with a correlation coefficient of 0.9997 and a detection limit of 3.68 mg/L.The recovery rate of spiked samples ranged from 93.8%to 105.8%,with relative standard deviations(RSD)ranging from 0.50%to 1.81%.

关 键 词：食糖 右旋糖酐 单克隆抗体 胶乳增强免疫比浊法 方法学 

分 类 号：TS201.3[轻工技术与工程—食品科学]