Addressing the problem of protein crystallization bottlenecks is a broadly regarded, but
complex, research topic. High-throughput screening techniques have been developed in the
recent decades to proceed with the investigations in this field. Some of these techniques are
regularly used to detect the best crystallizing formulations and form crystals that are suitable
for characterization. However, although slow crystallization processes can produce regular (or
large) crystals, a faster growth rate is aimed for. Videlicet, gaining a certain control over the
nucleation (or crystal growth) rate of proteins would be a breakthrough in modern science.
In this work, we focus on developing a low-concentration crystallizing solution and on
the design of a novel device to crystallize lysozyme from the above solution within an airdepleted
micro-batch environment. It was, de facto, observed, during this project, that the
heterogeneous crystallization of lysozyme in standard laboratories and from its conventionally
formulated solutions, hardly, would lead to a real understanding of this process. Videlicet, the
effect of the solid substrate topographies (or chemistries) on the lysozyme heterogeneous
nucleation rate might be altered also by interfering solution factors. Hence, the introduction of a
controlled crystallization environment, including surfaces bearing highly controlled features,
and the formulation of a low-salt precipitating solution were necessary.
This would also mean that, although, so far, a variety of surface effects on the
crystallization of conventionally formulated protein solutions have been observed, a far better
characterization of protein heterogeneous nucleation, and crystal growth, could be attained.
Videlicet, reducing the local effect of interfering, or uncontrollable, factors, almost certainly,
would lead to a better definition, and control, of the protein nucleation dynamics. Also, for the
same reason, it is pivotal that substrates with strictly controlled chemistries (or topographies)
are employed. Indeed, gaining a real understanding of the criteria that govern the protein
heterogeneous nucleation would be the ultimate scope of any work in this research field.