Potentiometric and spectroscopic studies of the reaction between trypsin and its inhibitors on chemically modified solid surfaces.

The potential of a titanium metal electrode modified with trypsin changes as a result of the complex formation reaction between trypsin and its inhibitor, aprotinin, dissolved in the solution. A similar potential change in the opposite direction occurs by the reaction between aprotinin-modified electrode and trypsin in the solution. The induced changes in both cases depend on the pH of the solution, showing the maximum change at pH = 9.5. The potentiometric response of the trypsin-modified electrode for the consecutive addition of aprotinin and proflavine proves that trypsin bound on the solid surfaces reacts with aprotinin much more strongly than with proflavine. This result is fully consistent with the spectroscopically observed behavior of a trypsin-modified quartz plate against these inhibitors. The surface coverage of trypsin on the quartz plate is also determined by a near-ultraviolet absorption measurement.