A chromatographic study of trypsin.

An analytical procedure for the resolution of trypsin and its derivatives would be useful in searching for active degradation products in tryptic autolysates as well as for other studies of the reactions of trypsin. Analytical procedures are frequently frustrated when applied to this enzyme because of the autolysis which occurs during the analysis. For this reason some previous analyses have been carried out on trypsin which had been stabilized by the conversion of the enzyme to acetyl trypsin (1). For preventing autolysis a more direct method, which would avoid complex side reactions which occur during acetylation (2) and which would allow the measurement of enzymic activity, might be based on the reversible inactivation of trypsin by urea (3). In general any resolving system, for example electrophoresis or chromatography, which maintained the trypsin in concentrated urea could give an analysis uncomplicated by autolysis and, upon dilution or removal of the urea, assays could be made of various fractions. This report describes the development of a technique for the chromatography of trypsin, in which autolysis is prevented by using urea in the eluting buffer.