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应用血清多肽SPG01、SPG03、SPG04的多抗(PcAb)酶联免疫吸附方法(ELISA),用于鼻咽癌放疗敏感性的检测。以碳二亚胺偶联法制备多肽免疫抗原,免疫动物制备兔血清多抗,用Protein A亲和层析法纯化多抗,选择3种多抗的工作滴度,应用多抗ELISA法检测63份鼻咽癌血清样本。紫外(UV)和聚丙烯酰胺凝胶电泳(SDS-PAGE)联合监测表明多肽偶联到BSA(牛血清白蛋白)上;动物免疫获得3种兔血清多克隆抗体各40 mL;纯化后的多抗纯度在90%以上,效价均在1∶100 000以上;3种多抗的检测滴度分别为1∶20 000,1∶100 000,1∶10 000;间接ELISA最低检测限为0.03μg/mL,相对标准偏差(RSD)为5.6%。t检验分析3种多肽区分鼻咽癌放疗敏感组与不敏感组,均呈显著差异(P<0.000 1)。ROC曲线表明,3种多肽检测鼻咽癌放疗敏感性的特异性在90%以上,灵敏度为73%~93%。3种多肽标志物均能显著区分鼻咽癌放疗敏感组与不敏感组,所应用的多抗ELISA方法检测鼻咽癌放疗敏感性特异性好,灵敏度高,可用于鼻咽癌个体化治疗的放疗敏感性辅助诊断。
Serum polypeptides SPG01, SPG03, SPG04 polyclonal antibody (PcAb) enzyme-linked immunosorbent assay (ELISA), for the detection of nasopharyngeal carcinoma radiosensitivity. The polyclonal antibody against rabbit polyclonal antibody was prepared by carbodiimide coupling method. The polyclonal antibody against rabbit serum was prepared by immunization. The polyclonal antibody was purified by Protein A affinity chromatography. The working titer of three polyclonal antibodies was selected. Nasopharyngeal carcinoma serum samples. The combined monitoring of UV and polyacrylamide gel electrophoresis (SDS-PAGE) showed that the polypeptide was conjugated to BSA (bovine serum albumin); the immunization of the three kinds of rabbit serum polyclonal antibody 40 mL each; Anti-purity was more than 90%, the titer was above 1: 100 000; the detection titers of three polyclonal antibodies were 1:20 000, 1: 100 000 and 1 1:10 000 respectively; the detection limit of indirect ELISA was 0.03 μg / mL with a relative standard deviation (RSD) of 5.6%. t-test analysis of three kinds of peptides to distinguish nasopharyngeal carcinoma radiotherapy sensitive group and non-sensitive group, were significantly different (P <0.000 1). The ROC curves showed that the specificity of the three peptides in detecting the radiosensitivity of nasopharyngeal carcinoma was over 90% and the sensitivity was 73% -93%. The three kinds of peptide markers can significantly distinguish nasopharyngeal carcinoma from radiosensitive group and non-sensitive group. The multi-anti-ELISA method applied to detect nasopharyngeal carcinoma radiosensitivity has good specificity and high sensitivity and can be used for individualized treatment of nasopharyngeal carcinoma Radiotherapy Sensitivity Assisted Diagnosis.