Inclusion Body Purification Service

Summary of Inclusion Body Purification Service

In the process of protein expression, researchers often use methods such as high temperature, high inducer concentration, and strong promoters to express the target protein at a high translation rate. These operations usually exhaust the bacterial protein quality control system, causing some inactive and insoluble protein molecules to aggregate in the cell to form inclusion bodies. Many documents have shown that 70%-80% of the recombinant proteins expressed in E. coli are inclusion bodies. The active proteins can be reversible from inclusion bodies in vitro, which indicates that inclusion bodies can be dynamic constructions (noncrystalline and amorphous structures) formed by the unbalanced equilibrium between aggregated proteins and soluble proteins in E. coli. Therefore, the use of inclusion bodies for membrane protein production is also considered an exciting method. The challenge at this time is no longer the purification of the recombinant protein, but how to dissolve and refold it into native and active protein. Of course, inclusion bodies are not only found in E. coli, they can also be formed in yeast, insect cells, and mammalian cells.

Inclusion bodies need extensive processing, including separation from cells, solubilization, refolding, and purification to produce bioactive proteins. Generally speaking, inclusion bodies recovered from cell lysates using low-speed centrifugation are heavily contaminated by E. coli cell wall and outer membrane components.

Inclusion Body Purification Service Provided by Creative BioMart Microbe

Creative BioMart Microbe has many years of experience in the purification of inclusion bodies, and has established an advanced technology platform that can meet the diverse needs of customers. The following is our experimental protocol designed to obtain native and soluble protein.

First, the inclusion bodies containing insoluble protein are separated from the cultured bacteria by homogenization, washing and centrifugation; then they are dissolved in denaturants (such as guanidine hydrochloride or urea) to refold them. It should be noted that certain reducing agents (such as DTT and GSSG) need to be added during this process to break the existing disulfide bonds and obtain monomeric peptide chains. We generally use detergent and low-concentration urea or guanidine chloride for selective extraction to dissolve protein pellets. Approximately 70% of the pellet protein can be dissolved through this basic operation. We have developed a solution to refold the protein correctly, and our researchers have accumulated a lot of expertise in protein recovery, which can ensure the native conformation and structural integrity of the target membrane protein.

Figure 1. Schematic inclusion body process (Humer, D.; Spadiut, O. 2018)

As an outstanding protein expression expert, Creative BioMart Microbe provides high-quality inclusion body purification service and techniques to help customers complete projects efficiently. Our advanced and unique technology platform can achieve more excellent purification efficiency and quality. The praise from customers all over the world is the proof of our quality service.

If you want to know more about this service, please feel free to contact us by phone or email, or you can consult through our inquiry form. We will ensure to provide you with efficient and high-quality experimental outsourcing services.

References

1. de Marco A.; et al. Bacterial inclusion bodies are industrially exploitable amyloids. FEMS Microbiology Reviews. 2019, 43.
2. Humer, D.; Spadiut, O. Wanted: more monitoring and control during inclusion body processing. World Journal of Microbiology and Biotechnology. 2018, 34:158.