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Nanobody-Based high-performance immunosorbent for selective beta 2-microglobulin purification from blood.
Acta Biomaterialia ( IF 9.7 ) Pub Date : 2020-02-22 , DOI: 10.1016/j.actbio.2020.02.028
Chundong Huang 1 , Jun Ren 1 , Fangling Ji 1 , Serge Muyldermans 2 , Lingyun Jia 1
Affiliation  

Removing β2-microglobulin (β2M) from blood circulation is considered to be the most effective method to delay the occurrence of dialysis-related amyloidosis (DRA). The ideal extracorporeal β2M removal system should be cost-effective, highly specific and having a high capacity. However, the traditional technologies based on size exclusion do not have an adequate specificity, and alternative immunosorbents have limited applications due to low capacity and their high cost. Nanobodies (Nbs), the smallest functional recombinant antibody fragments, offer several advantages to overcome these obstacles. In this study, an anti-β2M Nb with a C-terminal thiol-tag was successfully prepared from E. coli for site-directed and oriented immobilization and usage as capture ligand in a β2M-selective immunosorbent. The prepared immunosorbent showed a high binding capacity of up to 7 mg β2M per mL resin, which is 17 times higher than that of previous studies using single-chain variable antibody fragments (scFv). Furthermore, an exceptional high specificity has been demonstrated as other human serum proteins were not adsorbed during dynamic adsorption experiments. About 80% of the original binding capacity of the immunosorbent was restored after four consecutive easy regenerations, whereas 90% of the original capacity was retained after 1-month storage of the resin. Moreover, the mathematical model fitted very well the in vitro perfusion. The results with this pioneering immunosorbent confirm its possible clinical application and is expected to reach the required clinical effect of immunoadsorption therapy. STATEMENT OF SIGNIFICANCE: Dialysis-related amyloidosis (DRA), associated with the accumulation of β2-microglobulin (β2M), is a serious complication of end-stage kidney disease. Removing β2M from blood circulation by extracorporeal blood purification is considered to be the most effective method to delay the occurrence of DRA. However, the existing methods are incapable to eliminate sufficient quantities of β2M from circulation, either because of lack of specificity, high cost or for low capacity. In this manuscript, we provide a practical and economic immunosorbent based on anti-β2M nanobody for DRA. The prepared immunosorbent was reusable and storable, and demonstrated high specificity and realized a high binding capacity of up to 7 mg β2M per mL resin, which is 17 times higher than that of the previous studies.

中文翻译:

基于纳米抗体的高性能免疫吸附剂,可从血液中选择性纯化β2-微球蛋白。

从血液循环中去除β2-微球蛋白(β2M)被认为是延迟透析相关淀粉样变性病(DRA)发生的最有效方法。理想的体外β2M去除系统应具有成本效益,高度特异性和高容量。然而,基于尺寸排阻的传统技术没有足够的特异性,并且替代的免疫吸附剂由于容量低和成本高而受到限制。纳米抗体(Nbs)是功能最小的重组抗体片段,具有克服这些障碍的多项优势。在这项研究中,从大肠杆菌成功制备了具有C末端硫醇标签的抗β2MNb,用于定点和定向固定,并用作β2M选择性免疫吸附剂中的捕获配体。所制备的免疫吸附剂显示出高达7 mgβ2M/ mL树脂的高结合能力,这是以前使用单链可变抗体片段(scFv)的研究的17倍。此外,由于在动态吸附实验过程中未吸附其他人血清蛋白,因此显示出异常高的特异性。连续四次轻松再生后,免疫吸附剂的原始结合能力恢复了约80%,而树脂储存1个月后,免疫吸附剂的结合能力恢复了90%。此外,该数学模型非常适合体外灌注。这种开创性的免疫吸附剂的结果证实了其可能的临床应用,并有望达到免疫吸附疗法所需的临床效果。重要性声明:透析相关的淀粉样变性病(DRA),与β2-微球蛋白(β2M)的积累有关,是晚期肾脏疾病的严重并发症。通过体外血液净化从血液循环中去除β2M被认为是延迟DRA发生的最有效方法。然而,由于缺乏特异性,高成本或低容量,现有方法不能从循环中消除足够量的β2M。在本文中,我们提供了一种基于抗β2M纳米抗体的DRA实用且经济的免疫吸附剂。制备的免疫吸附剂可重复使用和存储,并显示出高特异性,并实现了高达7 mgβ2M/ mL树脂的高结合能力,这是先前研究的17倍。与β2-微球蛋白(β2M)的积累有关的是晚期肾病的严重并发症。通过体外血液净化从血液循环中去除β2M被认为是延迟DRA发生的最有效方法。然而,由于缺乏特异性,高成本或低容量,现有方法不能从循环中消除足够量的β2M。在本文中,我们提供了一种基于抗β2M纳米抗体的DRA实用且经济的免疫吸附剂。制备的免疫吸附剂可重复使用和存储,并显示出高特异性,并实现了高达7 mgβ2M/ mL树脂的高结合能力,这是先前研究的17倍。与β2-微球蛋白(β2M)的积累有关的是晚期肾病的严重并发症。通过体外血液净化从血液循环中去除β2M被认为是延迟DRA发生的最有效方法。然而,由于缺乏特异性,高成本或低容量,现有方法不能从循环中消除足够量的β2M。在本文中,我们提供了一种基于抗β2M纳米抗体的DRA实用且经济的免疫吸附剂。制备的免疫吸附剂可重复使用和存储,并显示出高特异性,并实现了高达7 mgβ2M/ mL树脂的高结合能力,这是先前研究的17倍。通过体外血液净化从血液循环中去除β2M被认为是延迟DRA发生的最有效方法。然而,由于缺乏特异性,高成本或低容量,现有方法不能从循环中消除足够量的β2M。在本文中,我们提供了一种基于抗β2M纳米抗体的DRA实用且经济的免疫吸附剂。制备的免疫吸附剂可重复使用和存储,并显示出高特异性,并实现了高达7 mgβ2M/ mL树脂的高结合能力,这是先前研究的17倍。通过体外血液净化从血液循环中去除β2M被认为是延迟DRA发生的最有效方法。然而,由于缺乏特异性,高成本或低容量,现有方法不能从循环中消除足够量的β2M。在本文中,我们提供了一种基于抗β2M纳米抗体的DRA实用且经济的免疫吸附剂。制备的免疫吸附剂可重复使用和存储,并显示出高特异性,并实现了高达7 mgβ2M/ mL树脂的高结合能力,这是先前研究的17倍。由于缺乏特异性,成本高或容量低,现有方法无法从循环中消除足够量的β2M。在本文中,我们提供了一种基于抗β2M纳米抗体的DRA实用且经济的免疫吸附剂。制备的免疫吸附剂可重复使用和存储,并显示出高特异性,并实现了高达7 mgβ2M/ mL树脂的高结合能力,这是先前研究的17倍。由于缺乏特异性,成本高或容量低,现有方法无法从循环中消除足够量的β2M。在本文中,我们提供了一种基于抗β2M纳米抗体的DRA实用且经济的免疫吸附剂。制备的免疫吸附剂可重复使用和存储,并显示出高特异性,并实现了高达7 mgβ2M/ mL树脂的高结合能力,这是先前研究的17倍。
更新日期:2020-02-22
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