Optimizing Macromolecule Crystallization: Creative Biostructure Service, Slides of Biology

Download Optimizing Macromolecule Crystallization: Creative Biostructure Service and more Slides Biology in PDF only on Docsity!

Optimization Screening

X-ray crystallography is the most successful methods for determining the

structure of macromolecules. However, obtaining crystals of

macromolecules is a very challenging task. The crystallization process can

be considered as a two-stage experiment, where the first stage determines

chemical and physical conditions under which the sample tends to

crystallize, while the second stage refines the parameters to produce

crystals suitable for analysis by X-ray diffraction. The quality and ease of X-

ray structure determination is directly correlated with the size and the

perfection of the crystalline samples. Initial crystals are frequently

microcrystals or clusters which often have unfavorable morphologies or

sizes, yielding poor diffraction intensities. It is therefore generally necessary

to improve upon these initial conditions in order to obtain better crystals of

sufficient quality for X-ray data collection.

Creative Biostructure has extensive experiences in refining crystallization conditions after initial screens. We use the grid screen or incomplete factorial approaches in the optimization process, in which sequential, incremental changes are made in the chemical parameters that influence crystallization, such as pH, ionic strength and reagent concentration, as well as physical parameters such as temperature, sample volume and overall methodology. Optimization can also entail the application of novel procedures and approaches that may enhance nucleation or crystal development. Our fully automated robotics allow us to complete combinatorial mixing from a wide range of stock solutions, followed by many possibilities for dispensing nano-volume screening experiments in the 96 well format.

Our crystallographers at Creative Biostructure perform optimization screening by careful manipulation and evaluation of biochemical, chemical, and physical variables, to produce crystals of sufficient size and quality for diffraction. We also provide guidance for our customers on how optimization might best be applied to crystal growth problems, and what parameters and factors might worth exploring to accelerate and achieve success. Please feel free to contact us for a detailed quote.

Thank you!

Excerpt from: https://www.creative-biostructure.com/optimization- screening-550.htm