Study On New Technology For Protein Array

In last several years, gene microarray has shown its great value in understanding the progression of human disease owing to the ability of analyzing the human gene sequence at the whole genome level. With the rise of proteomics, protein microarray has also played an important role in the research of proteomics by virtue of its high-throughout, high-specificity as well as high sensitivity. Moreover, impeled by proteomics, protein microarry technology develops quickly. Among protein microarray techniques, labeling detection technique has been applied abroadly owning to its high automation and high sensitivity. Recently, there have been a rising number of reports on microarray label-free detection published, showing that it has become an important direction for biological analysis due to its inherent merits of simple operational procedures and avoidance of the disturbance from either conjugated labels or management of radioactive materials. In recently years, protein microarray techniques are more and more frequently employed in the systematic research of proteomics, the discovery of disease biomarkers as well as drug screening etc, showing their great value in the research of biological researchIn this paper, two protein microarray detection techniques were investigated respectively, including labeling method and label-free method. In the labeling method, the agarose stamper was firstly integrated with silver enhancement method and applied to the fabrication and detection of the protein array. At the beginning, the homogeneity of the spots on the protein array and quantitative transferring capability of the agarose stamper as well as the activities of transferred proteins were checked by the fluorescent method. Then direct and sandwich reaction strategies were employed to perform quantitative analysis using the protein array based on this conjugation technique, respectively. The results demonstrated that the proteins can be transferred to the slide quantitatively and homogeneously (CV <10%). Moreover, the detection limit of 0.33fmol for direct direction and 0.13fmol for sandwich detection were obtained. Finally, in order to evaluate the effective number of repetitive stamping using the patterned agarose stamper, two indexes were introduced, and we found that seven protein arrays can be prepared in one time without intermediate re-inking. Owning to its merits of inexpensive and convenience, this integration technique has its promising to be a set of cost-effective, versatile and sensitive protein array fabrication and detection technique to be employed in the research of clinical diagnostic as well as proteomics. In the second experiment, Electrochemical impedance spectroscopy (EIS) combined with a gold electrodes array was developed to detect multiple antibody-antigen interactions. Hepatitis B surface antigen (HBsAg) as a model sample was employed to evaluate the characteristics of the biosensor. The array was...