Protein crystallization can be defined as the process involved in obtaining ordered and well-stacked lattices for complex macromolecules. Although protein crystallization is a science, it’s also viewed as an art as it requires interactions of certain variables for 100% success. 

Protein crystallization is employed to aid scientific investigations and experiments using mediums such as x-ray diffraction crystallography. Using this framework, researchers can accurately determine the molecular interactions by understanding the 3-dimensional protein blueprint. 

However, the relevance of protein crystallization doesn’t end there. For context, this technique also aids the formation of proteins devoid of other proteinous impurities or biological particles. 

In this guide, we’ll examine the process incorporated by protein production services to execute protein crystallization accurately. The subsequent lines will also examine the importance of protein crystallization to scientific research. 

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Major Steps to Ensure Protein Crystallization

For protein crystallization, the average protein production service adopts the following steps:

• Select an Appropriate Solvent

Although there are many solvents, picking a variation with the right amount of solubility is important. Additionally, researchers consider how safe the solvent is to use and how it’ll affect the energy and stability levels of the targeted protein or peptide. 

• Reach Supersaturation

Scientists have to heat the soluble peptide or protein to achieve supersaturation. The solution they get from this procedure will be supersaturated. 

• Crystallize Solution By Controlling Calcium Pump Proteins (CPPs)

After reaching supersaturation, researchers will cool off the supersaturated solution. This can be achieved via evaporation, reaction, or antisolvent addition. After solubility reduction, crystals will form and grow at a certain threshold. 

Although pure crystal products will grow in size, motifs with increased free energy will form non-crystal structures. To prevent the latter from happening, careful CPP control is required.

• Allow System to Achieve Equilibrium

Upon CPP control, scientists allow the system to achieve equilibrium. This step is taken as it’ll yield more product solids. To conclude the entire protein crystallization process, drying and filtering of the product are executed. 

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Importance of Protein Crystallization

Protein crystallization, since its inception, has reiterated its importance in the following areas:

• Medicinal Chemistry

Protein crystallization advancements and massive changes in x-ray crystallography have given room for the detection of thousands of 3D (three-dimensional) proteins. Understanding the interactions in these proteins is made possible by identifying the design of these macromolecules and their binding areas. 

The information from these interactions is crucial in understanding allosteric activator and protein indicator levels. It also helps determine active sites — an important aspect in developing medications with higher efficacy. 

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• Formation of Protein-based Therapeutics

Besides using protein crystallization to create single protein crystals, the scientific world has adopted this technique to develop protein-based therapeutics. The goal of these creations is to increase the development of stable and pure protein crystals with the right particle structure and a balanced particle size distribution. 

With this in view, protein crystallization is the most budget-friendly alternative for a protein production CRO to efficiently separate and purify macromolecules. This technique will also aid choral molecule isolation. 

• Effective Drug Delivery

Protein-based therapeutics gotten from protein crystallization can be administered to patients via injections. Due to the high amount of protein integrated into a single dose, protein crystallization comes in handy to dilute these proteins to relegate aggregation and viscolation issues to the background. 

For effective drug delivery via injections, protein crystallization helps to control the size of proteins to ensure they can pass through an infusion or injection needle. This, in turn, curbs the development of side effects. 

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Challenges Associated With Protein Crystallization

Although it seems the above steps are all needed for seamless protein crystallization, this process relies on numerous biochemical, chemical, and physical factors. 

Thus, protein crystallization remains an arduous task as it involves getting the mix of chemicals right, understanding the reactions of various proteins in multiple phases, and the ability to decipher nucleation and crystal growth in these proteins. 

To counter these difficulties, robots that denote precision have been developed. Although this revolutionary technological equipment is great, smaller custom recombinant protein production services can’t afford the expenses accompanying these add-ons. 

Major Takeaway

After examining protein crystallization closely, we’ve concluded that while it’s great for scientific research and treatment development, it’s still an art and science under construction. 

With crystal growth characterized by thousands of molecular attributes segmented into varying structures (primary, secondary, and tertiary), protein crystallization remains an endeavor guided by empirical understanding rather than clear-cut theory.