Karlsson Göran, Winge Stefan
Plasma R&D, Octapharma AB, SE-112 75, Stockholm, Sweden.
Protein Expr Purif. 2002 Oct;26(1):106-10. doi: 10.1016/s1046-5928(02)00510-7.
Human native antithrombin (AT) can be converted to a partially denaturated form of AT, known as latent AT (L-AT). This latent form of AT has been shown to exhibit strong antiangiogenic activity and also to suppress tumor growth in mice models. In the present work, a method is presented which induces the conversion of native AT to L-AT, using incubation at 60 degrees C, for 16 h, with 0.9 M ammonium sulfate, in 5mM Hepes buffer, pH 7.4, giving a recovery of more than 70%. L-AT was determined by integration of the low heparin affinity peak when analyzed by the affinity chromatography method. Native polyacrylamide gel electrophoresis was used to show that the preparation contained no aggregates. Hydrophobic interaction chromatography was also used for the separation of AT and L-AT.
人天然抗凝血酶（AT）可转化为部分变性的AT形式，即潜在抗凝血酶（L-AT）。这种潜在形式的AT已被证明具有很强的抗血管生成活性，并且在小鼠模型中还能抑制肿瘤生长。在本研究中，提出了一种方法，即在5mM Hepes缓冲液（pH 7.4）中，于60℃下用0.9M硫酸铵孵育16小时，诱导天然AT转化为L-AT，回收率超过70%。通过亲和色谱法分析时，通过对低肝素亲和力峰进行积分来测定L-AT。使用天然聚丙烯酰胺凝胶电泳来表明制剂中不含聚集体。疏水相互作用色谱法也用于分离AT和L-AT。
