固相基质表面von Willebrand Factor抗体的固定化研究

以丝素蛋白膜和聚乳酸膜为基质,采用辉光放电等离子体处理技术和共价交联的方法对vonWillebrand factor(vWf)抗体进行了固定化研究。利用抗体过剩法和酶联免疫实验对固定化效率进行了评价,固定化抗体的活性采用体外凝血时间(APTT,TT和PT)测定进行检测。结果显示固定化效率最高可以达到23.88%,酶联免疫实验均显示阳性,APTT、TT项有明显延长,有的还超出了仪器的测量范围,由此说明,通过这种方法可以有效地对vWf抗体进行共价固定化。本研究拓宽了抗体固定化技术的应用范围,为血管性假血友病的早期快速诊断仪器或试剂盒的研制打下技术基础,同时为其它凝血因子抗体相关的生物材料研究与开发提供一种新思路。

Immobilization of von Willebrand Factor Antibody on Solid Host Membranes

This study aimed at immobilizing the antibodies on the surfaces of the solid host membranes in order to improve the property of the biomaterial. The von Willebrand factor (vWf)antibodies were immobilized on the surface of Bombyx mori silk fibroin and PLA(Poly Lactic Acid) membrane by NH3 plasma treatment followed by covalent cross-linking reaction. The immobilization efficiency was evaluated by two methods including the antibody surplus and enzyme-linked immunosorbent assay (ELISA). The in vitro antithrombogenicity representing the activity of immobilized vWf antibody was determined by the method of Activated Partial Thromboplastin Time (APTT), Prothrombin Time (PT) and Thrombin Time (TT) test. The results demonstrate that the vWf antibodies are immobilized on silk fibroin and PLA membranes in an efficient way with the efficiency of antibody surplus up to 23.88% and ELISA reaction is positive. APTT and TT exceeded the upper limits distinctly, but the value of PT did not change noticeably. The in vitro antithrombogenicity represented the is activity-retaining form of antibodies. These results extend the application of antibody immobilization technique and provide a new idea about the design of biomaterials relating to the coagulation factors.