Technique of modifying the glass surface for production of protein microchips

Protein microarray consists of a support (usually glass or polymer) and immobilized proteins (ferments, antibodies, etc.). Being one of the most widespread and accessible support material, glass still has a significant disadvantage: its surface doesn’t contain functional groups capable of bonding with proteins. Therefore, the glass surface needs to be modified in order to immobilize capture proteins.

The aim of this work was to develop a technique of chemical modification of the glass surface for fabrication of protein microarrays. In our study we varied the following experimental parameters: solvent for 3-aminopropyltriethoxysilane, time of silanization reaction, concentration of glutaraldehyde, composition of a buffer for printing the target protein (allophycocyanin-labelled human anti-IgE) on the modified surface. The efficiency of protein molecule immobilization was quantified using the intensity of spot fluorescence. As a result, it was shown that there was no influence of the solvent for 3-aminopropyl-triethoxysilane on the immobilization efficiency of the target protein on the modified surface, and the optimal time for the silanization reaction was 60 minutes. It was also shown that the optimal concentration of glutaraldehyde was 2.5 % (v/v) and the optimal composition of the printing buffer was phosphate-buffered saline with the addition of 4 % (v/v) glycerol.

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