High-resolution optical interferometry was used to investigate the
effects of forced solution convection on
the crystal growth kinetics of the model protein lysozyme. Most
experiments were conducted with 99.99%
pure protein solutions. To study impurity effects, ∼1% of
lysozyme dimer (covalently bound) was added in
some cases. We show that the unsteady kinetics, corresponding to
bunching of growth steps, can be
characterized by the Fourier components of time traces of the growth
rate. Specific Fourier spectra are uniquely
determined by the solution conditions (composition, temperature, and
flow rate) and the growth layer source
activity. We found that the average step velocity and growth rate
increase by ∼10% with increasing flow
rate, as a result of the enhanced solute supply to the interface.
More importantly, faster convective transport
results in lower fluctuation amplitudes. This observation supports
our rationale for system-dependent effects
of transport on the structural perfection of protein crystals. We
also found that solution flow rates >500
μm/s result in stronger fluctuations while the average growth rate is
decreased. This can lead to growth
cessation at low supersaturations. With the intentionally
contaminated solutions, these undesirable phenomena
occurred at about half the flow rates required in pure solutions.
Thus, we conclude that they are due to
enhanced convective supply of impurities that are incorporated into the
crystal during growth. Furthermore,
we found that the impurity effects are reduced at higher crystal growth
rates. Since the exposure time of
terraces is inversely proportional to the growth rate, this observation
suggests that the increased kinetics
instability results from impurity adsorption on the interface.
Finally, we provide evidence relating earlier
observations of “slow protein crystal growth kinetics” to step
bunch formation in response to nonsteady step
generation.