Using an RNA aptamer probe for flow cytometry detection of CD30-expressing lymphoma cells.
Using an RNA aptamer probe for flow cytometry detection of CD30-expressing lymphoma cells.
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DOI：
10.1038/labinvest.2009.113
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发表时间：
2009-12
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--
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适体是由短的寡核苷酸组成的小分子配体，其以高亲和力结合靶标。与抗体相反，作为合成寡核苷酸，适体具有较低的生产成本并且不引起抗原反应。因此，适体是疾病诊断和治疗的潜在试剂。在这项研究中，我们验证了荧光标记的RNA适体，这已被报道为在溶液中特异性结合小鼠CD 30蛋白，人CD 30蛋白识别完整的细胞。用培养的表达高水平CD 30的间变性大细胞淋巴瘤和霍奇金淋巴瘤细胞测试适体探针。流式细胞术和荧光显微镜显示适体探针在低浓度（0.3 nM）下与表达CD 30的淋巴瘤细胞特异性和敏感性结合。对来自健康供体的多个细胞系和核细胞进行的研究证实，CD 30适体和抗CD 30抗体（标准临床探针）识别同一组细胞。还显示了使用CD 30适体探针和抗体的流式细胞术分析的潜在应用。总之，开发的CD 30适体探针可以作为抗体的替代和/或补充，在诊断表达CD 30的淋巴瘤。
Aptamers are small molecular ligands composed of short oligonucleotides that bind targets with high affinity. In contrast to antibodies, as synthetic oligonucleotides, aptamers have lower production costs and elicit no antigenic reactions. Therefore, aptamers are potential agents for disease diagnosis and treatment. In this study, we validate a fluorescently labeled RNA aptamer, which has been reported to bind specifically to mouse CD30 proteins in solution, for human CD30 protein recognition on intact cells. The aptamer probe was tested with cultured anaplastic large cell lymphoma and Hodgkin′s lymphoma cells that express high levels of CD30. Flow cytometry and fluorescence microscopy showed specific and sensitive binding of the aptamer probe to CD30-expressing lymphoma cells at low concentrations (0.3 nM). Studies performed on multiple cell lines and nuclear cells from healthy donors confirmed that the CD30 aptamer and anti-CD30 antibody, the standard clinical probe, recognized the same set of cells. The potential application of multicolor flow cytometry analysis using the CD30 aptamer probe and antibodies was also shown. In conclusion, the developed CD30 aptamer probe could act as a replacement and/or a supplement for antibodies in the diagnosis of the CD30-expressing lymphomas.
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