title: A microfluidic, high throughput protein crystal growth method for microgravity.
abstract: The attenuation of sedimentation and convection in microgravity can sometimes decrease irregularities formed during macromolecular crystal growth. Current terrestrial protein crystal growth (PCG) capabilities are very different than those used during the Shuttle era and that are currently on the International Space Station (ISS). The focus of this experiment was to demonstrate the use of a commercial off-the-shelf, high throughput, PCG method in microgravity. Using Protein BioSolutions' microfluidic Plug Maker™/CrystalCard™ system, we tested the ability to grow crystals of the regulator of glucose metabolism and adipogenesis: peroxisome proliferator-activated receptor gamma (apo-hPPAR-γ LBD), as well as several PCG standards. Overall, we sent 25 CrystalCards™ to the ISS, containing ~10,000 individual microgravity PCG experiments in a 3U NanoRacks NanoLab (1U = 10(3) cm.). After 70 days on the ISS, our samples were returned with 16 of 25 (64%) microgravity cards having crystals, compared to 12 of 25 (48%) of the ground controls. Encouragingly, there were more apo-hPPAR-γ LBD crystals in the microgravity PCG cards than the 1g controls. These positive results hope to introduce the use of the PCG standard of low sample volume and large experimental density to the microgravity environment and provide new opportunities for macromolecular samples that may crystallize poorly in standard laboratories.
authors.affiliation: Houston Methodist Research Institute, Department of Genomic Medicine, Houston, Texas, United States of America.
authors.forename: Carl W
authors.initials: CW
authors.lastname: Carruthers
authors.forename: Cory
authors.initials: C
authors.lastname: Gerdts
authors.forename: Michael D
authors.initials: MD
authors.lastname: Johnson
authors.forename: Paul
authors.initials: P
authors.lastname: Webb
mesh_terms: D003460:Crystallization; D044085:Microfluidics / Q000379:methods*; D011506:Proteins / Q000737:chemistry*; D014893:Weightlessness
chemical_list: D011506:Proteins