摘要
本文报道用抗ＰＡＩ－１单克隆抗体（ＭｃＡｂ）亲和层析建立了纯化ＰＡＩ－１的简便方法。经免疫亲和层析，ＳｅｐｈａｃｒｙｌＳ２００凝胶过滤，从ＨｅｐＧ２细胞培液中分离到糖基化和非糖基化两种形式的ＰＡＩ－１，回收率为８４％，ＰＡＩ－１比活性６．１×１０４ＩＵ／ｍｇ。糖基化ＰＡＩ－１分子量为５０ｋＤ，比活性５．８×１０４ＩＵ／ｍｇ。非糖基化ＰＡＩ－１分子量４３ｋＤ，占总ＰＡＩ－１３０％，仍具有ＰＡＩ－１活性。用ＣｏｎＡ－Ｓｅｐｈａｒｏｓｅ亲和层析进一步纯化得到ＳＤＳ－ＰＡＧＥ纯的糖基化ＰＡＩ－１。
A simplified procedure for the purification of PAI-1 from HepG2 cell based on immunoaffinity chromatography of anti-PAI-1 monoclonal antibodies(McAbs) was described.After immunoaffinity chromatography and Sephacryl S200 gel filtration, glycoprotein and non-glycoprotein forms of PAI-1 were seperated from HepG2 cell. The yield and the specific activity of putified PAI-1 were 84% and 6.1×104IU/mg, respectively. The MW of glycoprotein form of PAI-1 was about 50kD. The specific activity was 5.8×104IU/mg. The M W of non-glycoprotein form was 43kD. The non-glycoprotein form of PAI-1 contained 30%of total PAI-1 and also had activity of PAI-1. Glycoprotein form of PAI-1 was further purified by ConA-Sepharose chromatography.
关键词
PAI-1
纤溶酶原
激活剂抑制物
PAI-1
Anti-PAI-1 McAb
Immunoaffinity chromatography
HepG_2 cell
