A Preliminary Study Of The Bladder Tumor Protein Chip

Objective:Use human serum IgG for the target detection protein. Preparing the simultaneously detection multi-target protein chips, and further reduce the detection limit,as the prophase test preparation of bladder cancer multi-target detection.Methods:On the nitrocellulose membrane according to coubling dilution simultaneously coating six kinds of antibodies, preparing protein chips. Use different dilution ratio of IgG (standard) as the detection protein antigen specifically crossing with antibodies coated on the nitrocellulose membrane, and then adding the Biotinlation goat anti-human antibody IgG of Optimum concentratio, finally hybridizing with streptavidin added the colloidal gold label. After completion of staining applying the chip scanner to analysis gray value of the optical signal, and protracted the standard curve of different concentration gradient of IgG in the samples to the gray value of optical signal in order to find the best coated condition, and then to further reduce the detection lower limit.Results:1.The protein chip could be hybridized specifically with the corresponding antigens, no cross reaction.2.The detection lower limit could be up to100ng/ml, with good linear and gradient relationship, sensitivity up to100ng/ml.3. Comparing with protein chip in the37℃incubator,biotin-streptavidin-protein chip method to detect more quickly,no more obviously diffrences in the detection lower limit,and it had good repeatability, better beneficial to the late clinical detection.4.Found that the protein microarray is a kind of fast, specificity, sensitive detection methods.Conclusion:The chip could be used to detect multiple antigen targets simultaneously, and it is an effective new method of antibody detection,compared with the traditional colloidal gold method,The detection lower limit could be up to100ng/ml.