Protocol for growing insulin crystals of uniform size

• Protocol for growing insulin crystals of a uniform size is formulated and tested experimentally.
• Simple model predicting time evolution of the crystal size and supersaturation is devised and tested.
• Crystal nucleation and growth are decoupled in a batch crystallization process.
• Number density of nucleated crystals is preset to avert crystal nutrition competition.
• Experimental conditions to prevent new nuclei formation and to avoid nuclei loss are established.

Guidelines for growing insulin crystals of a uniform size are formulated and tested experimentally. A simple theoretical model based on the balance of matter predicts the time evolution of the crystal size and supersaturation. The time dependence of the size is checked experimentally. The experimental approach decouples crystal nucleation and growth processes according to the classical nucleation-growth-separation principle. The desired nucleation course was predetermined by carefully selecting the most suitable crystal nucleation parameters. Crystalline substance dispersity is predetermined during the nucleation stage of a batch crystallization process. To avert nutrition competition during the crystal growth stage, the number density of nucleated crystals is preset to be optimal.