The Establishment Of DAB2IP Detection Method And Its Application In Different TCM Syndromes Of Lung Cancer

Objective:To establish a DAB2IP fluorescence immunochromatographic test kit for detecting DAB2IP molecule, and explored the relating between the DAB2IP and TCM syndrome types of lung cancer, so as to guide the TCM treatment.Method:B lymphocyte hybridoma technique was applied to obtaina hybridoma cell line secreting anti-human DAB2IP antibody. The subclass of the mAb was identified through Mouse monoclonal antibody subtype detection kit.Ascites were induced to produce the monoclonal antibody from BALB/C mouse, and were purified with affinity chromatography method. Western blot was used to analyse the specific of the antibody.DAB2IP fluorescence immunochromatographic test kit was established by the fluorescence immunity chromatography; fluorescence immunochromatographic test kit used to detect the expression of DAB2IP in different TCM syndromes of lung cancer.Result:Two hybridoma named2A4、2G8were obtained which were able stably secreting monoclonal antibody against human DAB2IP molecule. The subclass of2A4、2G8were both mouse IgG1through mouse monoclonal antibody subclass detection kit. Ascites were induced to produce the monoclonal Antibody and5.0ml ascites was obtained from each BALB/C mouse on average. The purified antibody run glue show its molecular weight is about150kD; the Western blot display2A4McAb could react specifically with protein, the specificity is good; ELISA method for measuring DAB2IP was successfully established. The detecting limit was l.Ong/ml, precision degree is6.08%, linear in the range of1.00～500.OOng/mL.It evidenced that the kit wag of high sensitivity、precision and accutacy.Using the kit to detect the expression of DAB2IP molecules in different clinical syndromes of lung cancer, showed that deficiency of both qi and Yin group and the other group differences were significant (P<0.05), has obvious statistical significance.Conclusion: Two anti-human DAB2IP monoclonal antibodies and DAB2IP fluorescence immunochromatographic test kit with independent intellectual property rights were successfully obtained. Trough semi quantitative analysis of DAB2IP content in serum, the kit provide experimental basis for its application in research and clinical, and provide a new thought and method for lung cancer targeted therapy. As the expression of DAB2IP molecule in lung cancer with deficiency of both qi and Yin group and the other groups have significant differences (P<0.05), it suggest that by combining basic and modern immunology,may make the TCM type more objective, more deeply.At last the research has a certain reference value for the study of TCM modernization.