Characterization of anti-podoplanin monoclonal antibodies: Critical epitopes for neutralizing the interaction between podoplanin and CLEC-2
Characterization of anti-podoplanin monoclonal antibodies: Critical epitopes for neutralizing the interaction between podoplanin and CLEC-2
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DOI：
10.1089/hyb.2008.0017
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发表时间：
2008-08-01
期刊：
影响因子：
--
通讯作者：
Kato, Yukinari
中科院分区：
文献类型：
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作者：
Ogasawara, Satoshi;Kaneko, Mika Kato;Kato, Yukinari
Podoplanin （Aggrus）是一种黏液型唾液糖蛋白，被认为是淋巴内皮和肿瘤起始细胞（TICs）的有用标记物。据报道，podoplanin和c型凝集素样受体-2 （clc -2）之间的相互作用对podoplanin诱导的血小板聚集和癌症转移至关重要。最近，已经产生了几种抗人类podoplanin抗体；然而，这些抗podoplanin抗体尚未被很好地表征。采用酶联免疫吸附法（ELISA）、Western blot和流式细胞术检测合成的足平面蛋白肽和重组足平面蛋白缺失突变体的5种抗足平面蛋白抗体（nz1、D2-40、AB3、18H5和兔多克隆抗体）。z -1的表位是血小板聚集刺激（PLAG）结构域-2/3；D2-40、AB3和18H5的表位为PLAG1/2。D2-40和AB3的表位非常相似，尽管18H5与D2-40和AB3不同。流式细胞分析显示，nz1部分抑制了podoplanin和CLEC-2之间的相互作用，而其他抗体则没有。总之，两种最常用的抗podoplanin抗体D2-40和AB3具有相似的特性，尽管有几项研究报告了差异。nz1中和podoplanin和CLEC-2之间的相互作用，这可能导致针对podoplanin依赖的癌症转移的治疗性抗体的发展。
Podoplanin (Aggrus) is a mucin-type sialoglycoprotein that is known as a useful marker for lymphatic endothelium and tumor-initiating cells (TICs). Interaction between podoplanin and C-type lectin-like receptor-2 (CLEC-2) is reported to be critical for podoplanin-induced platelet aggregation and cancer metastasis. Recently, several anti-human podoplanin antibodies have been created; however, these anti-podoplanin antibodies have not been well characterized. Five anti-podoplanin antibodies (NZ-1, D2-40, AB3, 18H5, and a rabbit polyclonal antibody) were investigated using ELISA, Western blot, and flow cytometry with synthesized podoplanin peptides and deletion mutants of recombinant podoplanin. The epitope of NZ-1 is platelet aggregation-stimulating (PLAG) domain-2/3; the epitope of D2-40, AB3, and 18H5 is PLAG1/2. The epitopes of D2-40 and AB3 are quite similar, although 18H5 is different from D2-40 and AB3. Using flow cytometric analysis, NZ-1 partially inhibited the interaction between podoplanin and CLEC-2, although other antibodies did not. In conclusion, the two most frequently used anti-podoplanin antibodies, D2-40 and AB3, have similar properties, although several studies have reported differences. NZ-1 neutralizes the interaction between podoplanin and CLEC-2, which may lead to the development of therapeutic antibodies against podoplanin-dependent cancer metastasis.