JP2010204130A5 - - Google Patents

Claims (17)

A method of immobilizing a protein / peptide on a chip by activating a chemically modified diamond / DLC (Diamond-like Carbon) chip with an activating reagent and then spotting the protein / peptide on the chip to perform a coupling reaction a is, after activation by an activating reagent, have rows vacuum drying process of 30 to 120 minutes as drying of the washing water to wash the chips and, after the coupling reaction, 2-4 at 30-45 ° C. A method for immobilizing a protein / peptide, characterized by drying for a period of time . 2. The protein / peptide immobilization method according to claim 1, wherein the washing water is MilliQ water. 3. The protein / peptide immobilization method according to claim 1, wherein dimethyl sulfoxide (DMSO) or polyethylene glycol (PEG) is used as a spotting additive. The protein / peptide immobilization method according to claim 3, wherein 20-40% DMSO or 20-40% PEG is used. The method for immobilizing a protein / peptide according to claim 4, wherein 30% DMSO or 30% PEG is used. 6. The protein / peptide immobilization method according to any one of claims 1 to 5 , wherein the activating reagent is a solution containing WSCD.HCl and NHS. The method for immobilizing a protein / peptide according to any one of claims 1 to 6 , wherein an unreacted active group is blocked after the coupling reaction. The protein / peptide immobilization method according to any one of claims 1 to 7 , wherein the protein / peptide is an antigen or an antibody. 9. The protein / peptide immobilization method according to claim 8 , wherein the antigen is an allergen. 10. The protein / peptide immobilization method according to claim 9, wherein the allergen is an allergen epitope or a peptide containing an allergen epitope. 11. The protein / peptide immobilization method according to claim 10 , wherein a chemically modified peptide is used as the allergen epitope or the peptide containing the allergen epitope. As a peptide comprising an allergen epitope, claim 10, characterized in that an N-terminal and / or C-terminal side of at least two or more amino acids added in the epitope-containing peptides of a peptide moiety which binds to MHC class II molecules The protein / peptide immobilization method described. The method for immobilizing a protein / peptide according to any one of claims 10 to 12 , wherein a protease-degrading peptide containing an allergen epitope is used as the peptide containing an allergen epitope. 14. The protein / peptide immobilization method according to claim 13 , wherein the protease-degrading peptide is a trypsin-degrading peptide. The method for immobilizing a protein / peptide according to any one of claims 10 to 14 , wherein a food allergen is used as the allergen. As a food allergen, one or more allergens selected from sodium caseinate, α-casein, β-casein, κ-casein, α-lactalbumin, β-lactoglobulin, ovomucoid, ovalbumin, and conalbumin are used. The method for immobilizing a protein / peptide according to claim 15 . A protein / peptide immobilization chip obtained by the protein / peptide immobilization method according to any one of claims 1 to 16 .