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Syniheis and HIV capture activity of novel materials

新材料的 Syniheis 和 HIV 捕获活性

基本信息

批准号：
08458291
负责人：
BABA Masanori
金额：
$ 1.34万
依托单位：
Kagoshima University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
1996
资助国家：
日本
起止时间：
1996 至 1997
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08458291/
关键词：
HIV Nanosphere Lectin Monnose Capture Prophylaxis

项目摘要

HIV-1 gp120 has been shown to strongly interact with some lectins. Thus, gp120 and HIV-1 virions are expected to be effectively captured by lectins if they are immobilized to certain materials. Polymeric particles are often used as a material for immobilization of biomolecules such as antibodies and enzymes. Among them, polystirene particles are quite useful, since monodispersed particles can be easily prepared. Polystirene nano-spheres (NSs) covered with poly (methacrylic acid) were prepared by copolymerization of stirene with poly (tert-butylmethacrylayte) macromonomer and acid hydrolysis. Resulting NSs (mean diameter : 400nm) were dispersed well in water. The immobilization of concanavalin A (Con A) was performed by using water soluble carbodiimide. The surface of the obtained NSs (Con A-NSs) was found to be fully covered with Con A.The interaction of Con A-NSs with HIV-1 was determined by the reduction of gp120 level and viral infectivity of HIV-1 suspensions after 60-min incubation at room tem-perature. Con A-NSs achieved a>3.3 log and a 2.2 log reduction of viral infectivity in HIV-1 (III_B strain) suspension at a concentration of 2 and 0.5 mg/ml, respectively. Whereas Con A-free nanospheres, which were not immobilized with Con A,achieved only a 0.29 log reduction at 0.5 mg/ml. Con A-NSs (2 mg/ml) could also reduce the gp 120 level off III_B and HE strains to < 7.1 and 5.5% of each control, respectively. The combination of Con A-NS treatment followed by filtration with a microporous membrane efficiently removed virion-free gp120 as well as infectious viral particles from HIV-1 suspension. Scanning electron microscopy demonstrated that HIV-1 virions were trapped on the surface of Con A-NSs. Thus, Con A-NSs can capture HIV-1 virions and gp120 with a high affinity and may have potential as an effective tool for the prevention of HIV-1 transmission.

HIV-1 gp 120与某些凝集素有很强的相互作用。因此，如果gp 120和HIV-1病毒粒子被固定到某些材料上，则预期它们将被凝集素有效地捕获。聚合物颗粒通常用作用于固定生物分子如抗体和酶的材料。其中，聚苯乙烯颗粒是非常有用的，因为可以容易地制备单分散颗粒。通过苯乙烯与聚甲基丙烯酸叔丁酯大分子单体共聚和酸水解制备了聚甲基丙烯酸包覆的聚苯乙烯纳米球。所得的NS（平均直径：400 nm）在水中分散良好。采用水溶性碳二亚胺固定化刀豆球蛋白A（ConA）。结果表明，所制备的NS（ConA-NS）表面完全被ConA所覆盖。在室温下孵育60 min后，HIV-1悬液中gp 120水平和病毒感染性均降低，从而确定了ConA-NS与HIV-1的相互作用。浓度为2和0.5 mg/ml的Con A-NS分别使HIV-1（III_B株）悬液中的病毒感染性降低>3.3 log和2.2 log。而未用Con A固定的不含Con A的纳米球在0.5 mg/ml下仅实现0.29 log减少。ConA-NS（2 mg/ml）也可使Ⅲ_B和HE株的gp 120水平分别降低至各自对照的7.1%和5.5%。Con A-NS处理后用微孔膜过滤的组合有效地从HIV-1悬浮液中去除了无病毒体的gp 120以及感染性病毒颗粒。扫描电子显微镜显示HIV-1病毒粒子被捕获在ConA-NS表面。因此，Con A-NS能够以高亲和力捕获HIV-1病毒体和gp 120，并且可能具有作为预防HIV-1传播的有效工具的潜力。