A NOVEL INSTRUMENT AND METHOD FOR AMINO ACID ANALYSIS

A NEW APPROACH WILL BE DEVELOPED FOR PERFORMING AMINO ACID ANALYSIS OF PROTEIN HYDROLYSATES. IT WILL EMPLOY A UNIQUE DERIVATIZING REAGENT THAT IS CAPABLE OF REACTING WITH ALL AMINO ACIDS IN PROTEIN HYDROLYSATES. THESE INCLUDE PROLINE,CYSTEINE, AND TRYPTOPHAN, AMINO ACIDS THAT ARE TYPICALLY DIFFICULT TO QUANTITATE. THE AMINO ACID ANALYZER WILL BE BASED ON A HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY (HPLC) SYSTEM, BUT WILL CONTAIN AN ADDITIONAL COMPONENT FOR SAMPLE CLEANUP PRIOR TO INJECTION ONTO THE HPLC COLUMN. THIS SHOULD ELIMINATE THE SPURIOUS PEAKS TYPICALLY FOUND IN PREDERIVATIZATION PROCEDURES. THE METHODOLOGY WILL PROVIDE PICOMOLE-LEVEL SENSITIVITY, SHORT RUN TIMES, AND ACCURATE, DEPENDABLE RESULTS. AN INSTRUMENT BASED ON THIS NEW TECHNOLOGY WILL BE USEFUL IN BIOTECHNOLOGY AND MEDICAL RESEARCH LABORATORIES WORKING WITH BOTH NATURAL AND RECOMBINANT PROTEINS. THE USE OF PROTEINS IS INCREASING DRAMATICALLY, PARTICULARLY IN THE SEARCH FOR NOVEL PEPTIDE VACCINES AND IN THE DETAILED STUDY OF ANTIGEN-ANTIBODY INTERACTIONS. PHASE I WILL IDENTIFY UNIQUE DERIVATIZING AGENTS, CONSTRUCT A BREADBOARD ANALYZER,AND ESTABLISH THE CONDITIONS FOR AMINO ACID ANALYSIS. PHASEII WILL FURTHER VALIDATE THE ANALYTICAL METHOD WITH A VARIETY OF PROTEIN AND PEPTIDE SAMPLES FROM VARIOUS SOURCES. A PROTOTYPE COMMERCIAL INSTRUMENT WILL ALSO BE CONSTRUCTED TO AUTOMATE THE ENTIRE PROCEDURE.