Practical Protein Chemistry - A. Darbre 1989

Microscale amino acid sequence analysis using a gas-phase peptide-protein sequencer
Discussion
Chemical aspects

It has recently been demonstrated that in the step of FITC coupling to the peptide amino group, anhydrous TEA vapor can be used instead of aqueous TEA vapor without any noticeable effect on step yields, analytical sensitivity, and Background levels (R. M. Hewick, M. W. Hunkapiller, unpublished data). The anhydrous base has been successfully used in the analysis of both short Peptides and Proteins. The amino acid background obtained in the sequencing of Myoglobin under completely anhydrous conditions was just as low as that observed with the conventional aqueous TEA solution. These results indicate that the appearance of the amino acid background in the gas-phase Sequencer is caused mainly not by peptide bond Hydrolysis, but by other Chemical Reactions, as was previously suggested for the spinning-cup Reactor system [1].



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