Nano seeding tools to generate nanometer size crystallization seeds

Abstract

A kit and a method for using the kit to generate nanoseeds from protein nanocrystals and aggregates is disclosed. The method comprises mixing a plurality of beads with a protein nanocrystal or aggregate, and agitating the mixture to generate the nanoseeds. Nanoseeds made by disclosed embodiments may be of a high quality, as evaluated by TEM, and can be used to produce high quality protein crystals. Additionally, spectroscopic techniques, such as UV fluorescence and/or brightfield microscopy can be used to identify aggregates suitable to produce nanoseeds.

We claim: 
     
         1 . A kit, comprising:
 a container selected from a tube or a flask; and   a plurality of beads, each bead independently having a bead diameter suitable to be effective to make seed crystals from at least one protein nanocrystal.   
     
     
         2 . The kit of  claim 1 , wherein each bead independently has a bead diameter of from 0.05 mm to 1.5 mm. 
     
     
         3 . The kit of  claim 1 , wherein each bead independently has a bead diameter of from 0.1 mm to 1 mm. 
     
     
         4 . The kit of  claim 2 , wherein each bead independently has a bead diameter of less than 0.5 mm. 
     
     
         5 . The kit of  claim 2 , wherein each bead independently has a bead diameter selected from 0.1 mm, 0.5 mm, or 1 mm. 
     
     
         6 . The kit of  claim 1 , wherein the plurality of beads comprises glass beads, metal beads, metal alloy beads, ceramic beads, PTFE beads, or a combination thereof. 
     
     
         7 . The kit of  claim 1 , wherein the plurality of beads comprises glass beads, stainless steel beads, or a combination thereof. 
     
     
         8 . The kit of  claim 1 , wherein the plurality of beads comprises beads having a mixtures of diameters. 
     
     
         9 . The kit of  claim 1 , wherein the plurality of beads comprises beads of the same diameter. 
     
     
         10 . The kit of  claim 1 , wherein the tube is a microcentrifuge tube. 
     
     
         11 . The kit of  claim 10 , wherein the microcentrifuge tube has a size of from 0.5 mL to 2 mL. 
     
     
         12 . The kit of  claim 11 , wherein the microcentrifuge tube is a 1.5 mL microcentrifuge tube. 
     
     
         13 . The kit of  claim 1 , wherein the plurality of beads comprises from 2 to 1000 beads. 
     
     
         14 . The kit of  claim 1 , wherein the plurality of beads comprises from 2 to 500 beads. 
     
     
         15 . The kit of  claim 1 , wherein the plurality of beads comprises from 2 to 100 beads. 
     
     
         16 . A kit, comprising:
 a microcentrifuge tube; and   from 2 to 50 beads, each bead independently having a bead diameter of from 0.1 mm to 1 mm.   
     
     
         17 . The kit of  claim 16 , wherein the beads are glass beads, stainless steel beads, or a combination thereof. 
     
     
         18 . The kit of  claim 16 , wherein the bead diameter is selected from 0.1 mm, 0.5 mm, 1 mm or a combination thereof. 
     
     
         19 . The kit of  claim 16 , wherein the microcentrifuge tube has a size of from 0.5 mL to 2 mL. 
     
     
         20 . A kit, comprising:
 a microcentrifuge tube having a size of from 0.5 mL to 2 mL; and   from 5 to 30 glass and/or stainless steel beads, each bead independently having a bead diameter selected from 0.1 mm, 0.5 mm, 1 mm.