DOUBLE COUPLING EDMAN CHEMISTRY FOR HIGH-SENSITIVITY AUTOMATED PROTEIN SEQUENCING

Capillary electrophoresis with laser-induced fluorescence detection (C E-LIF) is a promising new method for the analysis of protein sequencin g products. It gives 10 zmol (1 zmol = 10(-21) mel) limits of detectio n (3 sigma) for fluorescein thiohydantoin (FTH) amino acids. We have d eveloped a separation for the (FTH)-amino acid products generated from 18 of the 20 coded amino acids. The extremely low volume requirement associated with CE-LIF makes it incompatible with commercial sequencer s, For this reason, we have also been developing a miniaturized sequen cer that can be more easily coupled to our detection system, Both the CE-LIF system and the miniaturized sequencer are described.