抗人IgG Fc片段及Fab段单抗的鉴定及应用

本实验采用木瓜酶水解,SPA柱亲合层析等手段得到人IgGFc段及Fab段,以Sigma抗人IgGfFc段和抗人IgG Fab段单抗为标准品,鉴定了细胞库中抗人IgG系列的部分细胞株,得到特异性分泌抗人IgG Fc段和抗人IgG Fab段单抗的细胞各一株。 在上述实验基础上,用抗人IgG Fc及抗人IgG Fab单抗分别制备了Sepharose4B亲合层析柱,提纯了酶解人IgG Fc、Fab片段,经ELISA法鉴定,相互之间无交叉反应。同时用此方法制备了人抗HBe Fab片段,并将该片段进行了过氧化物酶标记,用来配制HBe ELISA诊断盒,证明其生物活性未受影响,而且消除了类风湿因子引起的HBe Ag假阳性现象。因抗HBe单抗来源困难,如采用HBe多抗制备ELISA试剂,本法将是提高质量的一个好方法。

Identification and Application of Monoclonal Antibodies against Human IgG Fc and Fab Fragments

20monoclonal antibodies against human IgG were obtained by cell fusion After Hydrolysized with papain and separated on affinity chromatography SPA column, human IgG Fc and Fab fragments were available With these as coating antigens and sigma monoclonal antibodies to human IgG Fc and Fab fragments as standard substances, McAbs were identified and two of them were specific to human IgG Fc fragment and human IgG Fab fragment respectively. On affinity chromatography sepharose 4B columns prepared with each of the two strains,human IgG Fc and Fab fragments were purified and immunoassayed to be pure by ELISA. HBe ELISA kit was produced with above-mentioned purified human anti-HBe Fab fragments labelled by pero-xidase. and demonstrated no effect on their biology activities. This method has eliminated HBeAg pseudopositive phenomenon caused by rheumatoid factor. For lack of anti-HBe MeAb source, the method to prepare ELISA reagent with polyclonal anti-HBe would be better in improving quality.