Monoclonal Antibodies 13A4 and AC133 Do Not Recognize the Canine Ortholog of Mouse and Human Stem Cell Antigen Prominin-1 (CD133).
Monoclonal Antibodies 13A4 and AC133 Do Not Recognize the Canine Ortholog of Mouse and Human Stem Cell Antigen Prominin-1 (CD133).
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DOI：
10.1371/journal.pone.0164079
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发表时间：
2016
期刊：
影响因子：
3.7
通讯作者：
Corbeil D
中科院分区：
文献类型：
--
作者：
Thamm K;Graupner S;Werner C;Huttner WB;Corbeil D
五跨膜糖蛋白 prominin-1 (CD133) 作为干细胞和癌症干细胞的细胞表面标记物广泛应用于医学。它为基于干细胞的再生疗法和肿瘤学开辟了新途径。该分子主要用于人类样本或小鼠模型，因此大多数生物工具（包括抗体）都针对人类和鼠 prominin-1。尽管 prominin-1 的一般结构（包括其膜拓扑）在整个动物界中是保守的，但其一级序列保守性很差。因此，尚不清楚抗人和小鼠 prominin-1 抗体是否与其他物种（尤其是狗）中的直系同源物发生交叉反应。鉴于越来越多的研究使用犬 prominin-1 作为抗原标记，回答这个问题势在必行。在这里，我们通过克隆犬 prominin-1 并使用其在 Madin-Darby 犬肾细胞中作为绿色荧光蛋白融合蛋白的过度表达来解决这个问题，以确定其与针对人或小鼠 prominin-1 的抗体的免疫反应性。我们使用免疫细胞化学、流式细胞术和免疫印迹技术，令人惊讶地发现没有跨物种免疫反应性。当抗 prominin-1 抗体用于种间研究时，这些结果在数据解释中引起了一些注意。
The pentaspan membrane glycoprotein prominin-1 (CD133) is widely used in medicine as a cell surface marker of stem and cancer stem cells. It has opened new avenues in stem cell-based regenerative therapy and oncology. This molecule is largely used with human samples or the mouse model, and consequently most biological tools including antibodies are directed against human and murine prominin-1. Although the general structure of prominin-1 including its membrane topology is conserved throughout the animal kingdom, its primary sequence is poorly conserved. Thus, it is unclear if anti-human and -mouse prominin-1 antibodies cross-react with their orthologs in other species, especially dog. Answering this issue is imperative in light of the growing number of studies using canine prominin-1 as an antigenic marker. Here, we address this issue by cloning the canine prominin-1 and use its overexpression as a green fluorescent protein fusion protein in Madin-Darby canine kidney cells to determine its immunoreactivity with antibodies against human or mouse prominin-1. We used immunocytochemistry, flow cytometry and immunoblotting techniques and surprisingly found no cross-species immunoreactivity. These results raise some caution in data interpretation when anti-prominin-1 antibodies are used in interspecies studies.
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影响因子：
14.9
作者：
Dobson L;Reményi I;Tusnády GE
通讯作者：
Tusnády GE
DOI：
10.1006/bbrc.2001.5271
发表时间：
2001-07-27
影响因子：
3.1
作者：
Corbeil, D;Fargeas, CA;Huttner, WB
通讯作者：
Huttner, WB
影响因子：
14.9
作者：
Goujon M;McWilliam H;Li W;Valentin F;Squizzato S;Paern J;Lopez R
通讯作者：
Lopez R
影响因子：
1.2
作者：
Fujimoto, Ayumi;Neo, Sakurako;Tsuchiya, Ryo
通讯作者：
Tsuchiya, Ryo
影响因子：
4.3
作者：
Fargeas, Christine A.;Karbanova, Jana;Corbeil, Denis
通讯作者：
Corbeil, Denis