Protein Crystallography techniques

New techniques are revolutionizing structural biology

New technological advances and techniques in protein crystallography have inspired a structural biology revolution, enabling researchers to tackle even more complex biological problems to benefit scientific research. ‘Integrative’ structural biology brings together multiple approaches to build a more comprehensive picture of dynamic processes at both a cellular and molecular level. Protein crystallography remains the go to technique for high resolution protein structure studies.

Sitting drop crystallization

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Sitting drop crystallization is a commonly used protein crystallography method. For sitting drop, mosquito's precise pipetting ensures accurate drop-on-drop placement in the center of sub-wells for perfectly automated imaging whilst avoiding drops wicking up the well walls.

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Sitting drop
vapor diffusion

Using mosquito xtal3® for setting up vapor diffusion techniques has distinct advantages in multiple user research laboratories and academic groups, as both set-ups are easily automated without the need for changes in instrument configuration.

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Microbatch (Under-oil)

The multi-aspirate, dispense and drop mixing capabilities of mosquito crystal are perfect for microbatch set up. These enable both protein and screen solutions to be aspirated consecutively within the same tip and then be pipetted directly through oil to ensure that screen and protein drops always combine.

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Micro-Seeding

Seeding or micro matrix seeding (MMS) is a well-established method used to improve the quality and reproducibility of protein crystals. mosquito's plate set-up time is less than 3 minutes to create multiple aspiration.

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Additive screening

mosquito's unique true positive displacement pipettes combined with its multi-aspirate function allows users to access volumes down to 10 nL. Not only does this save on precious ligands/compounds, the contact dispensing ensures precise placement within the crystallization drops.

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Crystal soaking

Soaking protein crystals with ligands is the fastest route to produce high-throughput structures for fragment based drug discovery. mosquito can be used to prepare small soaking drops of DMSO based fragment libraries directly from stock plates. Small volumes of this diluted solution can then be introduced to the drop containing the crystal to save on time and ligand stock used.

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Hanging drop crystallization

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mosquito can automatically ‘mirror’ drops of crystal screen on a plate seal, so that when the seal is inverted over the plate, the drops match the screen wells. Precision placement and accurate pipetting mean that drop volumes can be miniaturized without the risk of the different components coinciding. Up to three drops can be created in each well of a 96-well microplate, allowing you to test up to 288 different conditions per plate.

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Bicelles

By maintaining a cold environment and pre-chilling the sample holder block it is possible to use mosquito crystal to accurately and easily pipette the bicelle solution, thereby facilitating the automation of screening for this lipidic based technique of membrane protein crystallization.

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Lipidic Cubic Phase (in-meso)

The Lipidic Cubic Phase (LCP) or in-meso technique for crystallizing membrane proteins has revolutionized structural studies of difficult targets such as G protein-coupled receptors (GCPRs). For handling Lipidic Cubic Phase (LCP) methods, mosquito® LCP combines four important aspects:

  • reduction of sample size down to nanolitre volumes
  • speed (about 4 minutes to complete a single 96-well LCP plate)
  • accuracy and repeatability
  • precise humidity control of the dispensing environment

mosquito LCP allows crystallography researchers to dispense Lipidic Cubic Phase (LCP) volumes as low as 30 nL, while automated calibration of syringe and pipette positioning ensures exceptionally precise drop-on-drop placement.

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Crystal screen optimization

Optimizing protein crystallization hits to improve diffraction is a challenging process, and identifying optimal conditions i.e. temperature, pH range, precipitants and concentration range of the buffer is particularly costly and time-consuming. The process involves the set-up of multiple studies to analyze each individual parameter so automating the preparation of complex gradients from stock solutions saves time, improves repeatability and increases the output of high resolution structures.

dragonfly crystal delivers accurate and repeatable results and its novel technology easily dispenses any volume into any well to streamline crystallization workflows and eliminate the risk of cross-contamination.

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Meet our Field Application Scientists

Our Field Application Scientists will help you to advance your research goals throughout the life of your instrument, optimizing for the applications you need and harnessing its full potential. As specialists in a range of disciplines from structural biology to genomics, the team ensures that the protocols for your applications are scientifically robust, giving you the confidence to pursue novel approaches.  Working closely with customers, Field Application Scientists eliminate bottlenecks and streamline workflows to enable successful research outcomes.

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