Moroi M, Goetze A, Dubay E, Wu C, Hasitz M, Jamieson G A
Thromb Res. 1982 Oct 1;28(1):103-14. doi: 10.1016/0049-3848(82)90038-x.
Platelet glycocalicin has been purified to homogeneity by a two step procedure involving affinity chromatography on WGA-Sepharose and then on thrombin-Sepharose using selective elution with heparin. The procedure is more rapid (3-4 days), more reproducible and gives about twice the yield (10 mg/40 units platelets) of the previous method (Okumura et al. J. Biol. Chem. 251, 5950-5955, 1976). The two preparations showed identical inhibition of aggregation of gel filtered platelets induced by thrombin and by ristocetin. Glycocalicin was cleaved in a controlled fashion by trypsin-Sepharose over 3hr to yield the macroglycopeptide and peptide "tail" fragments and there was no apparent further degradation with 18 hrs digestion.
血小板糖萼蛋白已通过两步法纯化至同质，该方法包括先在麦胚凝集素-琼脂糖凝胶上进行亲和层析，然后在凝血酶-琼脂糖凝胶上进行亲和层析，使用肝素进行选择性洗脱。该方法更快（3 - 4天），更具可重复性，产量约为之前方法（奥村等人，《生物化学杂志》251, 5950 - 5955, 1976）的两倍（10毫克/40单位血小板）。两种制剂对凝血酶和瑞斯托霉素诱导的凝胶过滤血小板聚集的抑制作用相同。糖萼蛋白在3小时内被胰蛋白酶-琼脂糖凝胶以可控方式切割，产生大糖肽和肽“尾”片段，18小时消化后没有明显的进一步降解。
