Study And Application Of Protein Microarray For Esophageal Cancer

Objective:Esophageal cancer (EC) is one of the most frequent killers among malignant diseases, ranking forth in China and seventh in the world, with five-year survival rate less than 10%. Screening of precancer and early stage cancer of esophagus is an important approach for reducing the incidence rate and death rate. Though balloon cytology and esophagoscopy approach are main ways now to screen esophageal precancerous individuals from high risk population, but they have many limits. So it is necessary to establish a new method to screen the esophageal precancerous patients from high-risk population in order to reducing the incidence rate and death rate of esophageal cancer.Methods:The16 SEREX identified antigens with statistical analysis were selected for preparing antigen microarray. The cDNA inserts of these 16 antigens were cloned into pET-30b (+) vector and expressed efficiently in E.coli. After purified by Ni-affinity column and Electro-elution, the recombinant fusion proteins were refolded in PBS buffer. Western blot revealed that the purity of these recombinant antigen proteins were over 90%.Then these 16 purified proteins were printed on the silylated slides by PC controlled stainless steel solid pins. With the antigen microarray, IgG and IgM autoantibodies to these 16 antigens were quantitatively detected in different sera groups, including sera from 241 EC patients, 35 patients with in situ carcinomas, 150 dysplasia patients, 234 normal donors, and sera from EC high incidence district donors, containing 323 perspective...