Agaylan Ashraf, Binder Daniel, Sauer Markus, Neuweiler Hannes, Meyer Oliver, Kiesewetter Holger, Salama Abdulgabar
CharitéCenter 14 of Tumor Medicine, Institute of Transfusion Medicine, Charité-Universitaetsmedizin Berlin, Berlin, Germany.
Cancer. 2007 Dec 1;110(11):2502-6. doi: 10.1002/cncr.23057.
Numerous assays have been described for the detection of p53 autoantibodies. These assays are highly specific with low sensitivity. In this report, the authors describe a highly sensitive and simple particle agglutination immunoassay using superparamagnetic particles for capturing p5 autoantibodies, p53 protein, and p53 protein-antibody complexes from large volumes of serum samples (2 mL).
Superparamagnetic particles were coated with different peptides spanning the entire p53 protein. These particles were incubated with serum samples from healthy blood donors (n=180), from patients without malignancies (n=27), and from patients with various forms of lung cancer (n=166). The particles were washed and placed into the reaction chamber of a gel card. After centrifugation, agglutination results were read visually. Positive reactions were defined by a layer of particles on top of the gel or agglutinated particles dispersed through the gel matrix.
Depending on the peptide used, p53 autoantibodies were detected in from 17.5% to 35% of the investigated patients with lung cancer. By using a commercially available enzyme-linked immunoadsorbent assay (ELISA) kit, p53 autoantibodies were detected in only 3% of those patients. P53 protein and p53 protein-antibody complexes were not detected in patients with lung cancer (n=20).
The newly developed assay was easy to perform and had sensitivity superior to that of the currently available p53 ELISAs.
已有多种检测p53自身抗体的方法被描述。这些方法具有高特异性但低灵敏度。在本报告中，作者描述了一种高灵敏度且简单的颗粒凝集免疫测定法，该方法使用超顺磁性颗粒从大量血清样本（2 mL）中捕获p5自身抗体、p53蛋白和p53蛋白 - 抗体复合物。
用覆盖整个p53蛋白的不同肽段包被超顺磁性颗粒。将这些颗粒与健康献血者（n = 180）、无恶性肿瘤患者（n =27）和各种形式肺癌患者（n = 166）的血清样本孵育。洗涤颗粒后将其放入凝胶卡的反应室中。离心后，肉眼读取凝集结果。阳性反应定义为凝胶顶部有一层颗粒或凝集颗粒分散在凝胶基质中。
根据所用肽段的不同，在17.5%至35%的肺癌患者中检测到p53自身抗体。使用市售酶联免疫吸附测定（ELISA）试剂盒，仅在3%的此类患者中检测到p53自身抗体。在肺癌患者（n = 20）中未检测到p53蛋白和p53蛋白 - 抗体复合物。
新开发的测定法易于操作，灵敏度优于目前可用的p53 ELISA。
