Affinity hemodialysis for antiviral therapy. I. Removal of HIV-1 from cell culture supernatants, plasma, and blood.

R. Tullis, R. Duffin, M. Zech, J. Ambrus
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引用次数: 38

Abstract

We tested an affinity hemodialysis technique designed to efficiently remove HIV and toxic viral proteins from blood. Miniature polyethersulfone hollow-fiber dialysis cartridges (200-500 nm pore) were packed with anti-HIV antibodies covalently coupled to agarose beads and sealed inside the cartridge. Cell culture fluids, plasma, or infected blood (7-15 ml) containing HIV-1 were circulated over the cartridge at 0.7-10 ml/min and the rate of removal of HIV measured by PCR and p24 ELISA. The technique removed up to 98% of HIV-1 particles from cell culture supernatants. Affinity hemodialysis also efficiently captured cultured HIV from human blood plasma (90%) and native HIV from infected blood (83% to 100%). Viral capture followed first-order kinetics (t(1/2) = 2.8 h). Variations in antibody type, matrix linkage (protein G versus direct coupling), bead pore size, and temperature of operation (25-37 degrees C) had only small effects. Although some binding was nonspecific, direct binding to the immobilized antibodies appeared to be the predominant mechanism.
亲和力血液透析用于抗病毒治疗。1 .从细胞培养上清、血浆和血液中去除HIV-1。
我们测试了一种亲和血液透析技术,旨在有效地去除血液中的HIV和有毒病毒蛋白。将抗hiv抗体与琼脂糖珠共价偶联,并密封在微型聚醚砜中空纤维透析盒(孔径200-500 nm)内。将含有HIV-1的细胞培养液、血浆或感染血液(7-15 ml)以0.7-10 ml/min的速度在药筒上循环,用PCR和p24 ELISA检测HIV的去除率。该技术从细胞培养上清液中去除高达98%的HIV-1颗粒。亲和血液透析也能有效地从人血浆中捕获培养的HIV(90%)和从感染血液中捕获原生HIV(83% - 100%)。病毒捕获遵循一级动力学(t(1/2) = 2.8 h)。抗体类型、基质连锁(蛋白G与直接偶联)、珠孔大小和操作温度(25-37℃)的变化只有很小的影响。虽然一些结合是非特异性的,但与固定抗体的直接结合似乎是主要的机制。
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