一种检测非小细胞肺癌患者血清中p53自身抗体的新方法

A Novel Method for Detecting p53 Autoantibodies in Sera of Patients with NSCLC

背景与目的多肽阵列是近年来发展起来的一种研究分子之间相互作用的方法。本研究拟建立多肽阵列检测非小细胞肺癌患者血清中p53自身抗体的方法,筛选p53自身抗体识别的抗原决定簇序列。方法利用点合成技术(SPOT synthesis technique)合成覆盖p53多肽链全部序列的多肽阵列;分别用ELISA和多肽阵列检测血清中的p53自身抗体,确定p53自身抗体识别的抗原决定簇序列。结果将包含p53多肽链全部序列的重叠肽段合成到硝酸纤维素膜上。多肽阵列检测30例非小细胞肺癌患者血清中的p53自身抗体,有7例为阳性;30例健康对照者均为阴性,与ELISA检测结果相同。P53自身抗体识别p53多肽链上某些共同的抗原表位。结论多肽阵列不仅可以用来检测肺癌患者血清中的自身抗体,还可以筛选这些自身抗体识别的抗原表位,为下一步研究自身抗体与非小细胞肺癌的早期诊断和预后的关系奠定基础。

Background and objective Serum autoantibody detection is useful means for the early diagnosis and prognosis of cancer. So our objective was to synthesize peptide array to analyse p53 autoantibodies in the sera of patients with non small cell lung cancer （NSCLC）. Methods Cellulose-bound overlapping peptides （12 mers） derived from p53 wild type protein were synthesized using SOPTs synthesis technique by an AutoSpot robot -ASP SL （Intavis, Germany）. ＂Ihe membrane was incubated with 1/400 dilutions ofpS3 monodonal antibody （Sc-53394） to establish a new approach to detect pS3 antibody, and the epitopes of the p53 monoclonal antibody is already known. We analysed the p53 autoantibodies from the sera of NSCLC and controls by peptide array and ELISA. Results We synthesized on cellulose membranes twelve-amino-acid overlapping peptides which included all of the sequences of the polypeptide chain of p53.The p53 autoantibody was positive in seven cases of thirty patients＇ sera with NSCLC and was negative in sera of the controls, with the same result of ELISA. Conclusion The peptide array could be applied not only to detect the autoantibodies in the sera of patients with lung cancer, but also to map the epitopes of the autoantibodies which might be useful for the early diagnosis and prognosis of cancer.