CD11d is a novel antigen on chicken leukocytes
CD11d is a novel antigen on chicken leukocytes
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DOI：
10.1016/j.jprot.2020.103876
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发表时间：
2020-04-15
影响因子：
3.3
通讯作者：
Hauck, Stefanie M.
中科院分区：
文献类型：
--
作者：
Deeg, Cornelia A.;Degroote, Roxane L.;Hauck, Stefanie M.
在生命科学中，抗体是用于识别、跟踪、定量和分离分子（主要是蛋白质）的最常用工具之一。然而，最近已经变得清楚的是，抗体在特异性和选择性方面常常不足，并且在许多情况下，靶蛋白甚至是未知的。当抗体的商业可用性稀缺时，例如用于靶向来自农场动物的蛋白质，研究人员面临额外的挑战：它们通常不得不依赖于交叉反应性抗体，而这些抗体的确切靶点、实际交叉反应性和所需应用的特征很差。在这项研究中，我们的目的是鉴定小鼠单克隆抗体8 F2的真正靶点，它是针对鸡PBMC产生的，并在研究中使用了几十年，而它的实际靶分子仍然未知。我们使用8 F2抗体在鸡PBMC中进行免疫沉淀，随后通过定量LC-MSMS鉴定其真正靶点为CD 11 d，这在鸡淋巴细胞中从未描述过。CD 11 d最丰富的相互作用物被鉴定为整合素β 2。因此，这种α整合素的存在在蛋白质水平上得到了明确的证明，并为进一步评估CD 11 d在鸡中的作用提供了第一个基础。数据可通过ProteomeXchange获得，标识符为PXD 017248。我们的研究确定了CD 11 d作为以前未表征的小鼠单克隆抗体8 F2的真正靶点，该抗体针对鸡外周血来源的单核细胞（PBMC）产生。因此，这是鸡中α整联蛋白的第一个成员，现在在蛋白质水平上清楚地鉴定出其存在。CD 11 d相互作用的额外鉴定提供了关于信号网络的整合素依赖性调节的信息，允许进一步的功能研究。
In life sciences, antibodies are among the most commonly used tools for identifying, tracking, quantifying and isolating molecules, mainly proteins. However, it has recently become clear that antibodies often fall short with respect to specificity and selectivity and in many cases target proteins are not even known. When commercial availability of antibodies is scarce, e.g. for targeting proteins from farm animals, researchers face additional challenges: they often have to rely on cross-reactive antibodies, which are poorly characterized for their exact target, their actual cross-reactivity and the desired application.In this study, we aimed at identifying the true target of mouse monoclonal antibody 8F2, which was generated against chicken PBMC and used for decades in research, while it's actual target molecule remained unknown. We used 8F2 antibody for immunoprecipitation in chicken PBMC and subsequently identified its true target as CD11d, which was never described in chicken lymphocytes before, by quantitative LC-MSMS. The most abundant interactor of CD11d was identified as integrin beta 2.The existence of this alpha integrin was therefore clearly proven on protein level and provides a first basis to further assess the role of CD11d in chickens in future studies.Data are available via ProteomeXchange with identifier PXD017248.Significance: Our studies determined CD11d as the true target of a previously uncharacterized mouse monoclonal antibody 8F2, generated against chicken peripheral blood derived mononuclear cells (PBMC). This is therefore now first member of alpha integrins in chickens, that existence was now clearly identified on protein level. The additional identification of CD11d interactors provides information on integrin-dependent regulation of signaling networks, allowing further functional studies.