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Application of antibody phage display to identify potential antigenic neural precursor cell proteins.
Journal of Biological Research-Thessaloniki ( IF 3.3 ) Pub Date : 2020-08-02 , DOI: 10.1186/s40709-020-00123-4
Ioannis Paspaltsis 1, 2 , Evangelia Kesidou 2 , Olga Touloumi 2 , Roza Lagoudaki 3 , Marina Boziki 2 , Martina Samiotaki 4 , Dimitra Dafou 5 , Theodoros Sklaviadis 1 , Nikolaos Grigoriadis 2
Affiliation  

The discovery of neural precursor cells (NPCs) and the concomitant intensive research in the field offer regenerative medicine novel approaches, enabling it to tackle conditions, such as neurodegenerative diseases. Transplantation of NPCs is nowadays considered a cutting-edge treatment for these conditions and many related clinical trials have been already completed or are still ongoing. However, little is known about the antigenicity of NPCs, with most studies addressing the question whether their antigenicity could lead to rejection of the transplanted cells. In this study we investigated the antigenic potential of syngeneic NPCs emulsion, upon subcutaneous (s.c.) administration to wild type C57BL/6 mice, following a standard immunization protocol. The whole IgG repertoire expressed upon immunization was cloned into a Fab phage display vector. From the created phage display library, Fab expressing clones interacting with NPCs lysate proteins were selected with the biopanning technique. The IgG Fab fragment from clone 65 proved to be reactive against antigens originating from NPCs lysates and/or whole brain lysate in diverse immunological assays. Using a standard immunization protocol to administer NPCs antigens, and applying the Fab fragment phage display technique, we were able to isolate at least a monoclonal IgG Fab fragment, which interacts with different mouse brain proteins. It is not clear whether such antibodies are produced in the host organisms, following NPCs transplantation.

中文翻译:

应用抗体噬菌体展示来鉴定潜在的抗原性神经前体细胞蛋白。

神经前体细胞 (NPC) 的发现以及该领域伴随的深入研究为再生医学提供了新的方法,使其能够应对神经退行性疾病等疾病。如今,NPC 的移植被认为是治疗这些疾病的前沿疗法,许多相关的临床试验已经完成或仍在进行中。然而,人们对 NPC 的抗原性知之甚少,大多数研究都解决了它们的抗原性是否会导致移植细胞排斥的问题。在这项研究中,我们按照标准免疫方案,在对野生型 C57BL/6 小鼠进行皮下 (sc) 给药后,研究了同系 NPC 乳液的抗原潜力。将免疫后表达的整个 IgG 库克隆到 Fab 噬菌体展示载体中。从创建的噬菌体展示库中,使用生物淘选技术选择与 NPC 裂解物蛋白相互作用的 Fab 表达克隆。来自克隆 65 的 IgG Fab 片段在多种免疫学测定中证明对源自 NPC 裂解物和/或全脑裂解物的抗原具有反应性。使用标准免疫方案来管理 NPC 抗原,并应用 Fab 片段噬菌体展示技术,我们能够分离出至少一个单克隆 IgG Fab 片段,该片段与不同的小鼠脑蛋白相互作用。目前尚不清楚这种抗体是否在 NPC 移植后在宿主生物体中产生。来自克隆 65 的 IgG Fab 片段在多种免疫学测定中证明对源自 NPC 裂解物和/或全脑裂解物的抗原具有反应性。使用标准免疫方案来管理 NPC 抗原,并应用 Fab 片段噬菌体展示技术,我们能够分离出至少一个单克隆 IgG Fab 片段,该片段与不同的小鼠脑蛋白相互作用。目前尚不清楚这种抗体是否在 NPC 移植后在宿主生物体中产生。来自克隆 65 的 IgG Fab 片段在多种免疫学测定中证明对源自 NPC 裂解物和/或全脑裂解物的抗原具有反应性。使用标准免疫方案管理 NPC 抗原,并应用 Fab 片段噬菌体展示技术,我们能够分离出至少一个单克隆 IgG Fab 片段,该片段与不同的小鼠脑蛋白相互作用。目前尚不清楚这种抗体是否在 NPC 移植后在宿主生物体中产生。
更新日期:2020-08-03
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