Question

In: Chemistry

How do we get crystals of protein by various methods?

How do we get crystals of protein by various methods?

Solutions

Expert Solution

Proteins are dissolved in an aqueous environment and sample solution until they reach a supersaturated state during the crystallisation process.There are various methods to get this supersaturated state.

 

  • Some of the methods include-

 

1. Vapour Diffusion - Droplets containing purified protein,          buffer, and precipitant equilibrate with a bigger reservoir        containing identical buffers and precipitants at higher            concentrations in this approach. The precipitant and              protein concentrations in the droplet of protein solution        are initially low, but as the drop and reservoir equilibrate,        the precipitant and protein concentrations in the drop           grow. Crystal growth happens in the droplet if the correct     crystallisation solutions are employed for a certain                 protein.

 

2. Microbatch- This technique usually involves immersing         of very small volume of protein droplet in oil.

 

3. Microdialysis- In this technique the sample is separated        from the crystallization reagent via way of means of a            semi-permeable membrane. The semi-permeable                    membrane lets in small molecules, together with salts,          additives, and different crystallization reagents to pass,        however prevents organic macromolecules from                    crossing the membrane.

 

4. Free-Interface Diffusion - This method brings collectively      protein and precipitation solutions with out premixing            them, however instead, injecting them via both facets of        a channel, permitting equilibrium via diffusion. The                  solutions come into touch in a reagent chamber, each at        their most concentrations, beginning spontaneous                  nucleation. As the system comes into equilibrium, the            extent of supersaturation decreases, favouring crystal            growth.


Various methods include -  

  1. Vapour diffusion.
  2. Microbatch.
  3. Microdialysis.
  4. Free-Interface distribution.

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