mosquito crystal: The benefits gained from using a high throughput microseeding technique for protein crystallisation

MMS application note, co-written with Dr Alexey Rak, Sanofi R&D, on the time- and cost-effectiveness of using microseeding in ambitious structural biology labs.

Protein crystallisation is a major bottleneck for protein structure determination, and it still remains the least standardised part of the protein structure determination process by X-ray crystallography.

Nucleation is the first and critical step to succeed with protein crystallisation. Crystal nuclei (seeds) can be transferred from drop to drop to increase success with the 3D growth of crystals. This seeding technique was mentioned in 1976 by Blundell and Johnson,1 where the standard microseeding technique was performed with a 10-fold dilution of crushed crystals.

In 1991, Enrico Stura introduced streak seeding and perfected macroseeding for setting up drops and the concept of using seeding to maneuver in the phase diagram without needing to calculate a phase diagram. This method revolutionised the protein crystallisation process.