摘要
本文将采用木瓜蛋白酶水解和ＳＰＡ－Ｓｅｐｈａｒｏｓｅ４Ｂ柱亲和层析等手段获得的人ＩｇＧＦｃ和Ｆａｂ，以抗人ＩｇＧＦｃ和抗人ＩｇＧＦａｂ单抗为参比品（Ｓｉｇｍａ）．鉴定了细胞库中抗人ＩｇＧ系列的部分细胞林，得到分泌特异性抗人ＩｇＧＦｅ和抗人ＩｇＧＦａｂ单抗的细胞各一株。在此基础上。应用抗人ＩｇＧＦｃ及抗人ＩｇＧＦａｂ单抗分别制备了Ｓｅｐｈａｒｏｓｅ４Ｂ亲和层析柱，纯化了酶解的人ＩｇＧＦｃ和Ｆａｂ。经ＥＬＩＳＡ法鉴定，相互间无交叉反应。同时用此方法还制备了人抗ＨＢｅＦａｂ，并将此Ｆａｂ标记过氧化物酶。配制ＨＢｅＥＬＩＳＡ诊断盒，证明其生物学活性未受影响，而且消除了类风湿因子引起的ＨＢｅＡｇ假阳性现象。
After hydrolyzed with papain and separated on SPA affinity chromatography column,human IgG Fc and Fab were available.With these as coating antigens and Sigma monoclonal antibodiee to human IgG Fo and Fab as reference substances, antihnman IgG McAbs were identified and two of them were specific for human IgG Fc and Fab, respectively.On Sepharos e 4B affinity chromatography columns prepared with each of the two McAbs,human IgG Fc and Fab were purified and immunoassayed to be pure by ELISA.HBe ELISA kit was produced with above-mentioned purified human anti-HBe Fab labed ed by peroxidas e and demonstrated no effect on it's biological activities.This method has eliminated HBeAg pseudopositive phenomenon caused by rheumatoid factor.
关键词
单克隆抗体
亲和层析
免疫球蛋白G
human IgG Fc and Fab
antibody,monoclenal
affinity chromatography
