Optimizing of the preparation condition of aldehyde glass slide for the production of proteinchip

An optimized method has been established to prepare and primarily evaluate protein-chip using glass slide.First the glass slide was washed and then different concentrations of3-Aminopropyltriethoxysilane(ATES)and glu taraldehyde was used to active the surface of the glass slide in different times.The optimized condition was de-cided by way of analyzing the kinetic relationship between the capacity of protein immobilization and concentrations of reagents and their during time of reaction.Series of10times diluted human IgG was printed on the surface of the glass.Cy3labeled second antibody was added on and after incubation and washing,the glass slide was scanned with laser confocal scanner.The dynamic range and the highest con centration that can be detected was defined through the strength of fluorescence of each concen tration.When the concentration of ATES and glutaraldehyde is4%and2.5%respectively and the treating time is30min and60min respectively the glass slide begin to have the highest capacity of immobilization of protein.The fluorescent signal have a good ki netic relationship with the con-cen tration of human IgG at the range between2mg /ml and2×4 -7 mg /ml.The highest concentration that can be de-tected is2×4 -8 mg /ml.