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。
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.
出处
《免疫学杂志》
CAS
CSCD
北大核心
1991年第2期114-117,共4页
Immunological Journal
