Use of high-performance liquid chromatography for preparing samples for microsequencing.

A method has been developed for the microsequencing of protein at subnanomole levels. The protein is carboxymethylated and freed of salts and reagents by reversed-phase chromatography prior to automated Edman degradation on a gas-phase sequencer. The carboxymethylated protein can also be fragmented chemically or enzymatically for further sequence analysis. The analytical techniques used to monitor the progress of the reactions all have picomole level sensitivity.