猪CD103单克隆抗体的制备与鉴定,Applied Microbiology and Biotechnology

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猪CD103单克隆抗体的制备与鉴定
Applied Microbiology and Biotechnology ( IF 3.9 ) Pub Date : 2022-05-23 , DOI: 10.1007/s00253-022-11950-x
Tao Zhang ₁ , Haoyuan Yu ₁ , Manita Aryal ₁ , Jing Yang ₁ , Maolin Li ₁ , Shuxian Li ₁ , Na Zhang ₁ , Han Shi ₁ , Baoyu Li ₁ , Guangliang Liu ₁ , Yuguang Fu ₁

Affiliation

摘要

树突状细胞 (DC) 在激活、调节和维持免疫反应中起重要作用。CD103+ DCs是DC亚群之一，主要在黏膜免疫反应中发挥作用。它们负责捕获抗原并将其携带到相关淋巴结以激活下游免疫反应。然而，关于 CD103+ DCs 在猪黏膜免疫反应中的功能的可用信息有限。本研究制备了两种针对猪 CD103 的单克隆抗体 (mAb)，并通过酶联免疫吸附试验 (ELISA)、间接免疫荧光试验 (IFA) 和流式细胞术对其应用进行了评估。生产的 mAb（7F3 和 9H3）均为 IgG1 亚型，轻链中有 κ 链。7F3 识别一个线性表位 (PDLRPRAQVYFSDLE)，而 9H3 识别另一个线性表位 (QILDEGQVLLGAVGA)。制备的mAbs可用于体内检测表达CD103分子的细胞，为两种mAbs提供了广泛的应用。综上所述，本研究成功制备了2个针对CD103蛋白的单克隆抗体，并在体内实验中显示出适用性，为猪黏膜免疫的研究提供依据。

关键点

• 制备针对猪CD103分子的单克隆抗体

• 可以分析CD103蛋白在不同细胞上的分布

• 探索猪黏膜免疫中的 CD103 ⁺ DCs 功能是可能的

"点击查看英文标题和摘要"

Preparation and identification of monoclonal antibodies against porcine CD103

Abstract

Dendritic cells (DCs) play an important role in activating, regulating, and maintaining the immune response. CD103+ DCs, one of the DC subpopulations, mainly function in the mucosal immune response. They are responsible for capturing and carrying antigens to the relevant lymph nodes to activate the downstream immune responses. However, there is limited available information regarding the function of CD103+ DCs in the porcine mucosal immune response. In this study, two monoclonal antibodies (mAbs) against porcine CD103 were prepared, and their applications were evaluated by enzyme-linked immunosorbent assay (ELISA), indirect immunofluorescence assay (IFA), and flow cytometry. The produced mAbs (7F3 and 9H3) were both IgG1 subtype with κ chains in the light chain. The 7F3 recognizes a linear epitope (PDLRPRAQVYFSDLE) while 9H3 recognizes another linear epitope (QILDEGQVLLGAVGA). The prepared mAbs could be used in vivo to detect the cells expressing CD103 molecules, giving wide applications of both mAbs. In conclusion, this study successfully prepared 2 mAbs against CD103 protein, and they showed applicability in vivo experiments, which will provide the basis for the study of porcine mucosal immunity.