A crystallization technique for obtaining large protein crystals with increased mechanical stability using agarose gel combined with a stirring technique

We developed a protein crystallization technique using a 0.0-2.0 w/v% agarose gel solution combined with a stirring technique for the purpose of controlling the crystal number in the gelled solutions. To confirm the stirring effect in the gelled solution, we investigated the nucleation probability and growth rate of the crystals produced using this method. The stirring operation by a rotary shaker affected the behavior of protein molecules in the gelled solution, and both a significant decrease in the nucleation rate and an enhancement of the crystal growth rate were achieved by the method. As a result, we concluded that the proposed technique, the stirring technique in a gel solution, was effective for generating protein crystals of sufficient and increased mechanical stability.