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A Microfluidic High Throughput Protein Crystal Growth Method for Microgravity

机译:用于微重力的微流体高通量蛋白质晶体生长方法

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摘要

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 = 103 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.
机译:微重力作用下沉降和对流的减弱有时可以减少大分子晶体生长过程中形成的不规则性。当前的陆地蛋白晶体生长(PCG)能力与航天飞机时代使用的以及现今国际空间站(ISS)的能力有很大不同。该实验的重点是演示在微重力中使用现成的,高通量的商用PCG方法的方法。使用Protein BioSolutions的微流体Plug Maker™/ CrystalCard™系统,我们测试了生长葡萄糖代谢和脂肪形成调节剂晶体的能力:过氧化物酶体增殖物激活受体γ(apo-hPPAR-γLBD),以及几种PCG标准。总体而言,我们向国际空间站发送了25张CrystalCards™,其中包含在3U NanoRacks NanoLab(1U = 10 3 cm。)中进行的约10,000个微重力PCG实验。在ISS上放置70天后,我们的样品将返回25张(64%)具有晶体的微重力卡中的16张,而地面对照中的25张(48%)则为12张。令人鼓舞的是,微重力PCG卡中的apo-hPPAR-γLBD晶体多于1g对照。这些积极的结果希望将低样品量和大实验密度的PCG标准引入微重力环境,并为可能在标准实验室中结晶不佳的大分子样品提供新的机会。

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