人纤维蛋白降解产物D-双聚体和E片段的纯化和鉴定

PURIFICATION AND CHARACTERIZATION OF CROSS LINKED FIBRIN FRAGMENT D-DIMER AND E FRAGMENT

D-双聚体和E片段均为交联型纤维蛋白的降解产物。根据它们的等电点的不同,建立了等电聚焦法纯化D-双聚体和E片段的方法。纯化的D-双聚体分子量约3.32×10^(-22)kg,含γ-γ双聚体片段(分子量1.36×10^(-22)kg)、β链片段(分子量7.30×10^(-23)kg)和α链片段(分子量2.49×10^(-23)kg)。Western Blot结果表明它可被抗D-双聚体单克隆抗体(McAb)识别。纯化的D-双聚体用ELISA检测,证实不含有E片段。纯化的E片段分子量约7.47×10^(-23)kg,还原后形成α链片段(分子量约1.03×10^(-23)kg)、β链片段(分子量6.14×10^(-24)kg)和γ链片段(分子量1.49×10^(-23)kg)3条肽链,可与抗E片段McAb进行免疫学反应。

The D-Dimer and E fragment are degradation products of cross-linked fibrin. Based on their different Pis, a simple isoelectric focusing procedure for the purification of D-Dimer and E fragment was established. The molecular weight of purified D-Dimer was 3.82×10-22kg. It is composed of a γ-γ dimer (molecular weight 1.86×10-22kg) αβ chain (molecular weight 7.80×10-23kg) and an a chain (molecular weight 2.49×10-23kg). Western Blot indicated that purified D-Dimer can be recognized by a monoclonal antibody against the D-Dimer. The purified D-Dimer contains no demonstrable E antigen comfirmed by ELISA. The molecular weight of E fragment was 7.47×10-23kg, that is composed of an a chain(ruolecular weight 1.03×10-23kg),αβ chain(molecular weight 6.14 ×10-24kg) and a γ chain (molecular weight 1.49×10-23kg). The purified E fragment reacted with monoclonal antibodies against E fragment.