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小鼠单克隆IgM抗体Fabμ片段的制备及其初步应用

Preparation and Application of Fabfμ Fragments from Mouse Monoclonal IgM Antibodies

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【作者】 鲁仲云周北平

【Author】 Lu Zhongyun & Zhou Beiping(Department of Immunology, Wuhan Institute of Biological Products, Ministry of Public Health, Wuhan 430060)

【机构】 卫生部武汉生物制品研究所免疫学研究室卫生部武汉生物制品研究所免疫学研究室 武汉 430060武汉 430060

【摘要】 在37℃和4℃下用胰蛋白酶消化小鼠单克隆IgM 4h和24h。SDS-聚丙烯酰胺凝胶电泳(SDS-PAGE)证实,37℃下制备的小片段分子量(Mr)近50000(Fab),但免疫活性较低;4℃下制备的小片段Mr为80000~110000(Fabμ),免疫活性与原IgM基本一致。选用在4℃下经胰酶消化IgM 24h制备的Fabμ与辣根过氧化物酶连接,其结合物用于双抗体夹心酶联免疫吸附试验,取得了满意的结果。

【Abstract】 This paper presents a method for preparation of small molecular active fragments from mouse monoclonal IgM antibodies. We have compared the influence of temperture on the splitting products at 37℃ and 4℃ by trypsin digestion. The fragments obtained at 37℃ had the Fab of molecular weight (Mr) approximate 50000 as determined by SDS-polycrylamide gel electrophoresis (PAGE) , its immunological activity is lower. But at 4℃, Mrs of the fragments are from 8 0000 to 110000(Fabn) , its immunoactivity is similar to the original IgM. The Fabu. (trypsin-treated IgM for 24h at 4℃ were labelled with horseradish peroxdase (HRP). The Fabμ·HRP conjugates have been used in sandwich-ELISA with satisfactory resutls.

【关键词】 小鼠IgM单克隆抗体Fabμ片段
【Key words】 Mouse IgMMonoclonal antibodiesFabμ fragments
  • 【文献出处】 免疫学杂志 ,Immunological Journal , 编辑部邮箱 ,1991年02期
  • 【被引频次】2
  • 【下载频次】60
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