Development Of Rapid Detection Reagent For Tumor Screening(Heat Shock Protein 90?)

Objectives: Heat shock protein 90 alpha(HSP90 alpha)is a kind of original stimulate cell induced by stress of stress protein,HSP90 alpha expression in tumor tissue of normal tissue expression level 2 to 10 times.In the positive cor relation between tumor malignant degree and the content in the blood.HSP90 alpha as a new tumor marker for lung cancer,gastric cancer,liver cancer,breast cancer,colorectal cancer,and other tumor clinical detection.HSP90 alpha levels in plasma corresponding patients changes,can real-time,accurately reflect the treatment effect,provide clinically objective basis for doctors with diagnosis,treatment and prognosis.This experiment for the protein system research,the establishment of heat shock protein 90 in human serum alpha(HSP90 alpha)fluorescence immune chromatography quantitative detection method.Methods: 1?Buy HSP90 alpha fusion protein,to antigen,concentration and purity identi fication of biological activity,immune mice,using the method of indirect enzy me-linked immunosorbent(ELISA)titer identification,cell fusion,monoclonal antibody preparation,single anti purity and subtype identification of protein purification process such as the preparation of high purity of HSP90 alpha mono clonal antibody and polyclonal antibody,for later use.2?Indirect the technology of fluorescence microshers immunochromatographicas say was used to research fluorescent microsphere activation,ensure optimal testing time,determine linearity range,precision,recovery experiments,testing clinical samples and that sort of thing in the fluorescence immunochromatograph icassay experiment.Results:A total of 16 strains screened monoclonal cell lines,after screening subtypes were selected 8 strains of positive cell lines,number3B2?4E3?4F4?5E2?7D1?7E4?8F2?8G4.In all the reaction conditions,it was determinedthat the optimal teaction time was 5 minutes,and in the best line range(0.39ng/m L-100ng/m L)precision;the sensibility is 0.0578ng/ml;quality control materials of high recovery(30ng/m L)Intra-assay cv was 7.46%;quality control materials of low recovery(10ng/m L)Intra-assay cv was 8.39%.With high,medium and low recovery of serum recovery was 100.56%,99.76%,94.1%,separately.Foreign kit(ELISA)for detection of 40 cancer patients clinical serum,the result showed that the corre lation is 0.9685.Conclusions: 1?Through the process of the preparation of monoclonal antibodies,successful 8 strains screened positive cell lines,and pick out a pair of mat ching antibodies used in the following experiment research.2?Ultrasonic technology in the process of activation of fluorescent microspheres can reduce gathered phenomenon 3?Establishing the method initially quantitativea fluorescence immunochromatographicassay for the heat shock protein 90?(HSP90?)in human serum have high performance and linear,clinic accordance rate also can satisfy the demand ofclinicquantitative detection,and will improvehopeful to applied to clinic apprroved.