Characterization of basigin monoclonal antibodies for receptor-mediated drug delivery to the brain
Characterization of basigin monoclonal antibodies for receptor-mediated drug delivery to the brain
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DOI：
10.1038/s41598-020-71286-2
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发表时间：
2020-09-03
影响因子：
4.6
通讯作者：
Nielsen, Morten Schallburg
中科院分区：
文献类型：
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作者：
Christensen, Sarah Christine;Krogh, Berit Olsen;Nielsen, Morten Schallburg
脑疾病生物治疗药物的脑吸收受到血脑屏障（BBB）的阻碍。血脑屏障选择性地调节大分子进入大脑的运输，从而维持大脑的内稳态。受体介导的胞吞作用（RMT）是一种将必需蛋白传递到脑实质的机制。在脑内皮细胞中表达的受体已被探索以双功能抗体形式通过血脑屏障运送治疗性抗体。在这项研究中，我们生成并鉴定了与basigin受体结合的单克隆抗体（mab），该受体最近被提出作为血脑屏障RMT的靶点。抗体结合特性，如亲和力，已被证明是胞吞能力和效率的重要因素。然而，关于basigin mAb特性对RMT影响的研究是有限的。在这里，我们描述了不同的basigin单克隆抗体与人脑内皮细胞结合并随后内化的能力。对单克隆抗体进行分析，以确定受体结合表位和亲和力是否影响受体介导的摄取效率。通过竞争性表位分组研究，将basigin单克隆抗体分为5个表位分组。来自三个表位箱的单克隆抗体通过结合特性及其在人脑内皮细胞中的高水平内化来判断RMT候选物所需的特性。
The brain uptake of biotherapeutics for brain diseases is hindered by the blood-brain barrier (BBB). The BBB selectively regulates the transport of large molecules into the brain and thereby maintains brain homeostasis. Receptor-mediated transcytosis (RMT) is one mechanism to deliver essential proteins into the brain parenchyma. Receptors expressed in the brain endothelial cells have been explored to ferry therapeutic antibodies across the BBB in bifunctional antibody formats. In this study, we generated and characterized monoclonal antibodies (mAbs) binding to the basigin receptor, which recently has been proposed as a target for RMT across the BBB. Antibody binding properties such as affinity have been demonstrated to be important factors for transcytosis capability and efficiency. Nevertheless, studies of basigin mAb properties' effect on RMT are limited. Here we characterize different basigin mAbs for their ability to associate with and subsequently internalize human brain endothelial cells. The mAbs were profiled to determine whether receptor binding epitope and affinity affected receptor-mediated uptake efficiency. By competitive epitope binning studies, basigin mAbs were categorized into five epitope bins. mAbs from three of the epitope bins demonstrated properties required for RMT candidates judged by binding characteristics and their superior level of internalization in human brain endothelial cells.