GROWTH OF PROTEIN CRYSTALS SUSPENDED IN A CLOSED-LOOP THERMOSIPHON

The quality of protein crystals often suffers from their growth at a liquid or solid surface. A novel solution growth method was developed to alleviate this problem. A growing crystal is suspended in a specially configured upflow of supersaturated nutrient, which is provided by the effect of fluid buoyancy in a closed loop thermosyphon. The flow rate and supersaturation are controlled by the temperature distribution in the thermosyphon, while contact of the crystal with the wall during growth is practically eliminated. The method was applied to the growth of lysozyme single crystals, with surprising results. While the orthorhombic form of lysozyme grew readily to the suspension limit of this particular apparatus (1.5 mm), the tetragonal form grew only to a maximum size less than 0.1 mm. Seed crystals of tetragonal lysozyme introduced into stagnant batch controls did not experience the growth cessation that the suspended crystals did. A likely cause of this growth cessation is the fluid shear forces on the suspended crystals.