Proteins adsorbed to a hydrophobic surface used for determination of proteolytic activity

Wettability of a thin layer of protein adsorbed to a hydrophobic surface is reduced after proteolytic digestion. Reduced wettability is demonstrated by condensation of water vapour on the surface. The condensation patterns of enzyme-treated and untreated protein layers give different light-scattering properties which can be observed by the naked eye. Based on these principles, a new simple and inexpensive method, thin layer enzyme assay (TEA), for determination of proteolytic activity, was developed. Fibrinogen, gammaglobulin (IgG), bovine serum albumin (BSA), haemoglobin, ovalbumin and gelatin were used as substrates. The proteolytic activity in 1 ng trypsin (EC 3.4.21.4) and in 1 ng pronase (EC 3.4.24.4) was reproducibly detected.