Aptamer directly evolved from live cells recognizes membrane bound immunoglobin heavy mu chain in Burkitt's lymphoma cells
Aptamer directly evolved from live cells recognizes membrane bound immunoglobin heavy mu chain in Burkitt's lymphoma cells
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DOI：
10.1074/mcp.m700026-mcp200
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发表时间：
2007-12-01
影响因子：
7
通讯作者：
Tan, Weihong
中科院分区：
文献类型：
--
作者：
Mallikaratchy, Prabodhika;Tang, Zhiwen;Tan, Weihong
肿瘤相关细胞膜蛋白靶点的鉴定对于理解肿瘤进展、开发新的诊断工具以及潜在地鉴定新的治疗靶点是重要的。在这里，我们提出了一种新的策略，用于确定蛋白质的表达水平改变，在病变细胞中使用细胞特异性适体。使用一个完整的活B细胞伯基特淋巴瘤细胞系（拉莫斯细胞）作为目标，我们选择了适体识别细胞膜蛋白具有高亲和力。在选择的对不同白血病细胞系显示不同识别模式的适体中，适体TD 05显示与拉莫斯细胞结合。通过化学修饰TD 05以与其在拉莫斯细胞上的靶标共价交联以使用链霉亲和素包被的磁珠捕获和富集靶受体，然后进行质谱分析，我们能够鉴定膜结合的免疫球蛋白重μ链作为TD 05适体的靶标。免疫球蛋白重μ链是B细胞抗原受体的主要组分，其在伯基特淋巴瘤细胞中表达。这项研究表明，这种两步策略，开发高质量的适体探针，然后鉴定其靶蛋白，可用于发现新的疾病相关的潜在标志物，从而提高肿瘤的诊断和治疗。基于适体的策略将使得能够有效地对疾病相关的生物标志物和其他感兴趣的分子进行分子阐明。
The identification of tumor related cell membrane protein targets is important in understanding tumor progression, the development of new diagnostic tools, and potentially for identifying new therapeutic targets. Here we present a novel strategy for identifying proteins that are altered in their expression levels in a diseased cell using cell specific aptamers. Using an intact viable B-cell Burkitt's lymphoma cell line (Ramos cells) as the target, we have selected aptamers that recognize cell membrane proteins with high affinity. Among the selected aptamers that showed different recognition patterns with different cell lines of leukemia, the aptamer TD05 showed binding with Ramos cells. By chemically modifying TD05 to covalently cross-link with its target on Ramos cells to capture and to enrich the target receptors using streptavidin coated magnetic beads followed by mass spectrometry, we were able to identify membrane bound immunoglobin heavy mu chain as the target for TD05 aptamer. Immunoglobin heavy mu chain is a major component of the B-cell antigen receptor, which is expressed in Burkitt's lymphoma cells. This study demonstrates that this two step strategy, the development of high quality aptamer probes and then the identification of their target proteins, can be used to discover new disease related potential markers and thus enhance tumor diagnosis and therapy. The aptamer based strategy will enable effective molecular elucidation of disease related biomarkers and other interesting molecules.