A system to remove b2-microglobulin from blood

描述(由申请人提供)： 这项建议的总体目标是开发一种专用于b2-微球蛋白(B2m)的血液解毒系统，b2-微球蛋白是一种淀粉样蛋白，与透析相关的淀粉样变性(DRA)有关。DRA是一种使人丧失能力的、可能致命的、不可避免的后果，是长期肾功能衰竭和当前医疗技术无法取代肾功能的所有方面的结果。肾脏缺乏B2M清除导致这种蛋白的血浆浓度升高，随后B2M-淀粉样蛋白通过尚未完全了解的机制在组织中沉积。尽管已经做了几次尝试来降低B2M的血浆浓度，但这种疾病的发病率和并发症仍然是一个重要的问题。有人建议研究一种体外免疫吸附方法，即使用固定化重组单链抗体片段来清除B2M。利用分子生物学和发酵技术将产生抗人B2M的单链可变区抗体片段(ScFv)，对其进行表征并将其固定在多孔载体上，以用于新的吸附器设计。该吸附器的有效性将在体外血液中进行测试，其生物相容性将在绵羊身上进行体外测试。拟议的研究结果可能导致以下结果：1)使用重组抗体片段免疫吸附器作为治疗方法来减缓DRA的进展，同时避免所需蛋白质的不分青红皂白地丢失；以及2)使用专门设计的免疫吸附器作为工具，帮助阐明可疑的中分子量可溶因素在终末期肾脏疾病发病率中的作用。使用DRA作为疾病模型来获得原理证明，将支持这项技术的量身定制的应用，以移除导致其他病理状态的靶分子。

DESCRIPTION (provided by applicant): The overall goal of this proposal is to develop a blood detoxification system that is specific for b2-microglobulin (b2m), an amyloidgenic protein that has been implicated in dialysis-related amyloidosis (DRA). DRA is an incapacitating, potentially fatal, and unavoidable consequence of long-term renal failure and the inability of current medical technology to replace all aspects of kidney function. The lack of b2m-clearance by the kidneys results in elevated plasma concentrations of this protein, followed by b2m-amyloid deposition in tissues via mechanisms that are not fully understood. Although several attempts have been made to reduce b2m plasma concentrations, the incidence and complications of this disease remain a significant problem. It is proposed to investigate an extracorporeal, immunoadsorptive approach that uses immobilized recombinant single-chain antibody fragments to remove b2m. Single-chain variable region antibody fragments (scFv) against human b2m will be produced using molecular biology and fermentation techniques, characterized and immobilized onto a porous support for use in a novel adsorber design. The efficacy of the adsorber will be tested in blood in vitro and its biocompatibility will be tested ex vivo in sheep. The results of the proposed research could potentially lead to the following: 1) use of a recombinant antibody fragment immunoadsorber as a therapy to slow the progression of DRA while avoiding the indiscriminate loss of needed proteins and 2) use of specifically designed immunoadsorbers as tool to help elucidate the role of suspected middle molecular weight soluble factors in the morbidity of end stage renal disease. Achieving proof of principle using DRA as a model disease would support the tailored application of this technology for the removal of target molecules that cause other pathologic states.