A numerical analysis of factor X activation in the presence of tissue factor--factor VIIa complex in a flow reactor.

A mathematical model has been developed to investigate previously obtained experimental findings relating to the activation of factor X by surface-bound tissue factor--factor VIIa (TF:VIIa) in a tubular flow reactor. In those experiments, factor X was perfused through a microcapillary tube over a range of flow (shear) conditions and the activated product, factor Xa, was measured at the outlet of the tube using a chromogenic assay. In the present study, the steady-state convection-diffusion equation with Michaelis-Menten kinetics used to describe the reaction at the wall was numerically integrated using an implicit method based on linear systems of ordinary differential equations. The results from the numerical analysis indicated that shear rate directly affects both Km and Vmax. Values of Km decreased from 151 to 16 nM as the shear rate increased from 25 to 2400 sec-1. Additionally, there was a twofold increase in Vmax from 1.4 to 3.0 pmol/cm2/min as the shear rate increased from 25 to 300 sec-1. These findings are in contrast with classical enzyme behavior and imply a direct effect of fluid flow on the kinetics of factor X activation.