中文摘要：

许多蛋白质二聚化或多聚化在调节其功能方面发挥重要作用,研究蛋白质的聚合有助于阐明相关的生物学过程.本文使用酶联免疫吸附法,对分子量11 k D的马传染性贫血病毒核壳蛋白（nucleocapsid protein 11 k D,NCp11）的聚合进行检测.首先利用亲和层析分别纯化了NCp11以及N-端或C-端融合有FLAG标签的NCp11.然后将NCp11包被于聚苯乙烯96孔板底,加入带FLAG标签的NCp11与之聚合,再依次加入抗FLAG抗体、辣根过氧化物酶标记的二抗及底物,反应终止后于450 nm波长下读取吸收值.结果表明,酶联免疫吸附法适用于NCp11聚合的检测,可对聚合的特异性、剂量依赖效应和影响因素等进行定量评价.利用该方法不仅能检测蛋白质的聚合,而且具有灵敏度高、特异性好、通量高、成本低和快速简便等优点,有望获得广泛应用.

英文摘要：

Dimerization or polymerization plays vital roles in the functional regulation of many proteins.Analysis of protein polymerization will help to understand many biological processes. In this study,enzyme-linked immunosorbent assay（ ELISA） was employed for the detection of protein polymerization.The 11 k D nucleocapsid protein（ NCp11） of equine infectious anemia virus was tagged with or without a FLAG motif at either N-or C-terminus. The recombinant protein was purified by affinity chromatography.ELISA was utilized to detect NCp11 polymerization. Native NCp11 was immobilized on the surface of micro-well plates. FLAG-tagged NCp11 was added for the polymerization. Anti-FLAG monoclonal antibody and horseradish peroxidase（ HRP）-labeled secondary antibody were used for ELISA.Influencing factors of NCp11 polymerization were evaluated. Quantitative detection of NCp11 polymerization appeared to be sensitive and specific,with high-throughput and cost effective potentials.