Besselink G A, Beugeling T, Beernink R, Poot A A, Van Aken W G, Bantjes A
Department of Chemical Technology, University of Twente, Enschede, Netherlands.
J Biomater Sci Polym Ed. 1995;7(7):551-62. doi: 10.1163/156856295x00472.
The adhesive proteins fibrinogen (FG) and fibronectin (FN) were immobilized to glycine-Sephadex G-10. The derivatized Sephadex G-10 gels were used to bind human blood platelets. For comparison, Gly-Arg-Gly-Asp-Ser-Pro(GRGDSP)-derivatized Gly-Sephadex G-10 was used. FG-, FN-, and GRGDSP-Gly-Sephadex G-10 each bound a substantial number of activated blood platelets (> or = 5 x 10(8) ml-1 gel) while non-activated platelets were not bound. Binding of ADP-treated blood platelets to the affinity adsorbents was dependent on the ADP-concentration which was used, reaching a near-maximal value at about 10 microM ADP. Platelet binding to the three types of affinity gels could be completely inhibited by dissolved GRGDSP as well as monoclonal anti-platelet glycoprotein IIb/IIIa (GPIIb/IIIa) antibody CLB-C17, which demonstrates that platelet binding specifically involves the fibrinogen binding site on GPIIb/IIIa. Platelet binding to all three affinity gels required free Ca2+ and Mg2+ ions: platelets binding in the absence of these divalent cations was considerably lower than platelet binding in buffer containing 2 mM Ca2+ and 1 mM Mg2+. Moreover, activated ethylenediamine-tetraacetate (EDTA)-treated platelets did not bind at all to the affinity gels. The finding that non-activated platelets did not bind to the affinity gels is thought to be related to both the high hydrophilicity of the Sephadex basic material and to the native state of the gel-bound fibrinogen and fibronectin.
将黏附蛋白纤维蛋白原（FG）和纤连蛋白（FN）固定到甘氨酸 - 葡聚糖凝胶G - 10上。将衍生化的葡聚糖凝胶G - 10用于结合人血血小板。作为对照，使用了甘氨酰 - 精氨酰 - 甘氨酰 - 天冬氨酰 - 丝氨酰 - 脯氨酸（GRGDSP）衍生化的甘氨酸 - 葡聚糖凝胶G - 10。FG -、FN - 和GRGDSP - 甘氨酸 - 葡聚糖凝胶G - 10各自结合了大量活化的血血小板（≥5×10⁸个/ml凝胶），而非活化的血小板则未结合。经ADP处理的血血小板与亲和吸附剂的结合取决于所用的ADP浓度，在约10μM ADP时达到接近最大值。血小板与这三种亲和凝胶的结合可被溶解的GRGDSP以及单克隆抗血小板糖蛋白IIb/IIIa（GPIIb/IIIa）抗体CLB - C17完全抑制，这表明血小板结合特别涉及GPIIb/IIIa上的纤维蛋白原结合位点。血小板与所有三种亲和凝胶的结合都需要游离的Ca²⁺和Mg²⁺离子：在没有这些二价阳离子的情况下血小板的结合明显低于在含有2 mM Ca²⁺和1 mM Mg²⁺的缓冲液中血小板的结合。此外，经活化的乙二胺四乙酸（EDTA）处理的血小板根本不与亲和凝胶结合。未活化的血小板不与亲和凝胶结合这一发现被认为与葡聚糖基础材料的高亲水性以及凝胶结合的纤维蛋白原和纤连蛋白的天然状态有关。
