Effective monitoring of protein reaction on glass plate surfaces by TOF-SIMS

Time-of-flight secondary ion mass spectrometry (TOF-SIMS) is capable of chemically visualizing proteins on insulated samples. Distribution of an immobilized probe protein, fluorescent-labeled protein A-immobilized on a glass plate, and that of a sample protein, immunogloblin G (IgG) in solution, reacting with protein A on the biosensor surface, were evaluated with TOF-SIMS (TFS-2100, Physical Electronics). TOF-SIMS spectra and images of the protein on the glass plates were obtained, and this “mutual information”, as defined by information theory, was employed to analyze the TOF-SIMS spectra of proteins. Fragment ions from protein A and IgG were distinguished by the mutual, reinforcing information and specific fragment ions to each protein were selected to obtain the TOF-SIMS image of the protein. It is evident from the TOF-SIMS images of each protein that protein A was immobilized on the substrate homogeneously and that the reaction between the immobilized protein A and IgG is not localized in this condition. Chemical images of the proteins by TOF-SIMS will contribute to a better understanding of the reaction on the biosensor surface, and thus will help the development of more sophisticated biosensors. In addition, the requisite chemical conditions as well as the interaction between the biosensor surface and the immobilized proteins were investigated by TOF-SIMS by means of sets of reinforcing, mutually supportive information.