Andoh K, Kubota T, Kobayashi N, Maekawa T
Thromb Res. 1986 Aug 1;43(3):275-86. doi: 10.1016/0049-3848(86)90147-7.
The apoprotein (AP) of tissue factor (TF) has been purified 72,000-fold to homogeneity from human placenta using acetone delipidation, sodium deoxycholate (DOC) extraction, Sephacryl S-300 column chromatography, preparative polyacrylamide-gel electrophoresis (PAGE) in DOC and tryptic digestion. The purified AP had an apparent molecular weight of 54,000 by sodium dodecyl sulfate/PAGE. A radioimmunoassay (RIA) for quantitation of the TF-AP using an antibody against this purified AP of TF was devised which was sensitive enough to measure as small a quantity as 100 pg/ml of TF-AP accurately with high reproducibility. In addition to TF clotting activity (TFA), the immunoreactive TF-AP (IR-TFR) in the homogenates of leukemic leukocytes from patients with acute non-lymphoid leukemia (ANLL) was determined using this RIA. In 30 patients with ANLL, the mean IR-TFR with standard deviation (SD) of 21 cases with DIC was 157.9 +/- 188.1 ng/10(8) cells, which was significantly higher than that (37.1 +/- 29.9 ng/10(8) cells) of 9 cases with no DIC during remission induction chemotherapy (p less than 0.01).
利用丙酮脱脂、脱氧胆酸钠(DOC)提取、Sephacryl S - 300柱色谱、在DOC中进行制备性聚丙烯酰胺凝胶电泳(PAGE)以及胰蛋白酶消化,已从人胎盘中将组织因子(TF)的载脂蛋白(AP)纯化至72000倍纯度且达到均一性。通过十二烷基硫酸钠/PAGE测定,纯化后的AP表观分子量为54000。设计了一种放射免疫测定法(RIA),用于定量TF - AP,该方法使用针对这种纯化的TF - AP的抗体,灵敏度足以准确测量低至100 pg/ml的TF - AP,且具有高重复性。除了TF凝血活性(TFA)外,还使用该RIA测定了急性非淋巴细胞白血病(ANLL)患者白血病白细胞匀浆中的免疫反应性TF - AP(IR - TFR)。在30例ANLL患者中,21例发生弥散性血管内凝血(DIC)患者的平均IR - TFR及其标准差(SD)为157.9±188.1 ng/10⁸细胞,显著高于9例在缓解诱导化疗期间未发生DIC患者的(37.1±29.9 ng/10⁸细胞)(p<0.01)。