镁离子对免疫胶体金检测中生物大分子相互作用的影响  

The effect of magnesium ion on interaction among biomacromolecules in immune colloidal gold assays

作　　者：王笑笑 林洪[1] 隋建新[1] 曹立民[1] WANG Xiao-xiao;LIN Hong;SUI Jian-xin;CAO Li-min(Food Safety Laboratory,Ocean University of China,Qingdao 26600)

机构地区：[1]中国海洋大学食品安全实验室,青岛266003

出　　处：《食品科技》2018年第7期298-306,共9页Food Science and Technology

基　　金：国家自然科学基金项目(31271864);山东省自主创新专项(2014ZZCX02703)

摘　　要：目的:探索镁离子对免疫胶体金检测中生物大分子相互作用的影响,为进一步阐释基质干扰机制、研发前处理新技术、提升检测效率提供参考依据。方法:通过斑点免疫金渗滤法分析了镁离子对免疫胶体金检测过程中主要生物大分子免疫反应的影响,并通过表面等离子共振技术进行动力学分析,进一步探索镁离子对大分子物质亲和活性的影响。结果:首次确定镁离子能够通过干扰金标抗体进而影响抗原抗体的特异性结合,进一步确定了镁离子的干扰使金标抗体的动力学常数明显不同,推测出镁离子影响了金标抗体的F^(ab)片段。结论:定性、定量分析了镁离子与免疫胶体金反应中主要生物大分子的相互作用,在此基础上明确了镁离子对金标抗体亲和活性的影响,该研究结果为食品质量安全领域胶体金快检技术的基质干扰研究、干扰控制消除及检测效能的改进提升,提供了进一步的理论依据和指导。Objective： To explore the effect of magnesium ion（Mg^（2＋）） on interaction among biomacromolecules in immune colloidal gold assays（ICA） and to provide the guidance for further explaining the mechanism of matrix interference, developing new technologies and improving the efficiency of detection. Methods： Based on dot immuno-gold filtration assay（DIGFA）, the effect of Mg^（2＋） on immunoreaction of gold-labelled antibody and antigen, gold-labelled antibody（GAB） and Ig G was analyzed. The effect of Mg^（2＋） on macromoleculars affinity was explored by SPR biosensor. Results： The general interference in ICA from Mg^（2＋） was found by DIGFA, Mg^（2＋） can affect the binding between GAB and antigen by impeding blinding site of GAB with confirmed for the first time. The kinetics constants between GAB and antigen were significantly different by SPR. Mg^（2＋） mainly interfered with the Fab fragment of the GAB was preliminarily inferred. Conclusion： Qualitative and quantitative analysis of the interaction of Mg^（2＋） and major biomolecules in the mainly reaction of immuno-colloidal gold, from which the effect of Mg^（2＋）on antibody affinity was clarified. The results of this study provided further theoretical reference and guidance for the research of interference mechanism, the improvement of the control and elimination of matrix interference and detection efficiency of colloidal gold rapid detection technology in food quality and safety field.

关 键 词：镁离子 相互作用 免疫胶体金技术 金标抗体 干扰 

分 类 号：R155.5[医药卫生—营养与食品卫生学] TS207.3[医药卫生—公共卫生与预防医学]