A protein is essentially an extended chain of linked amino acids arranged sequentially. This is called primary structure. In the end, though, proteins fold into extremely intricate three-dimensional conformations where different segments are attracted to or repellent from one another. The various portions of a protein molecule that fold into a complicated three-dimensional structure are designated by secondary, tertiary, or quaternary structures. In fact, the 3D structure of any proteins is crucial to maintain the its proper function.
There are various ways of obtaining functional and pure protein:
- One strategy is to isolate a peptide or protein by delicately lysing the tissues hypnotically. PurMa Biologics has decades of expertise in extracting 3D structure of various proteins for activity assay and other downstream applications which need restoration of the native structure of the protein. This is also an essential step in generating Monoclonal Antibodies. We have all the tools to anticipate and calculate the surface exposed epitopes of the proteins in order to generate an Monoclonal antibody which detects the native (3D) structure of the protein. Antibodies made against the linear sequence of the amino acids of a protein often fail to detect the protein inside the intact cells, such as immunohistochemistry.
- A second and more straight forward and at the same time risky methodology is to use other organelles such as bacteria (Ecoli) . in this procedure the cDNA of a gene which represents the coding sequence of a protein is cloned in an expression vector. Bacterial expression of a protein is handier for the small proteins and those with no transmembrane domains. In these cases, the expressed protein is soluble. The bigger proteins and the peptides with one or more membrane domains are accumulated inside the inclusion body of the bacteria and therefore is insoluble. . .
How Are Purification and Expression of Proteins Done in the Laboratory?
Protein Expression
In 1982, when the polymerase chain reaction (PCR) was discovered to allow for quick and controlled DNA duplication. Since then, scientists around the globe have been able to amplify any length of DNA and provide the cDNA for the expression of recombinant proteins in laboratory. PurMa Biologics with three decades of expertise in gene cloning as well as protein expression can provide great help to your research.
Protein Purification
During the expression of protein, other biological components will be mixed with the desired protein. PurMa Biologics has the latest technology in purification of your expressed proteins using the tag already cloned in the cDNA. With PurMa Biologics’s help, the pure extract will only contains your desired and expected protein free of additional biological material.
The Protein Expression and Purification Bottleneck
With the rise in popularity of PCR and its offspring (qPCR, RT-PCR, and RT-qPCR), research facilities have been looking for methods to expedite gene cloning, protein expression and purification. Once the lab overcome this step, there will be expedites the protein expression and purification much fastermove through their experiments as quickly as they can.
Optimization results in automation
By thoroughly analyzing the sequence of coding sequences and making sure of being in the right frame with the cloned tag ( such as GST, MBP, His or HA ), automation will occur and that is the answer to the bottleneck.
Automation in PCR protocol execution enables labs to finish experiments much faster, which frees up time for more lab experiments to be completed, examined, and published.
In PurMa Biologics, the Gene Cloning as well as protein expression and purification are done by experts with decades of experience in biochemical purification and troubleshooting.
Fully Automated Laboratory for Protein Expression and Purification
Even though protein expression and purification are now more flexible, dependable, and quick, thanks to automation, every lab can benefit from implementing Total Laboratory Automation (TLA) due to ongoing advancements in automation.
Many laboratories have already succeeded in protein expression purification. The time it takes to go from the first experimental design to the finished results has been reduced. Long-term TLA costs are significantly lower than those of conventional laboratory techniques, even though the initial cost might be high.
Some challenges for TLA still need to be solved, like the requirement for sufficient lab space and the possibility of downtime if automated machinery malfunctions. However, thanks to microchip technology’s ever-increasing dependability and ever-diminishing size, every obstacle is manageable for any lab that aspires to Total Laboratory Automation.
Contact PurMa Biologics to arrange a consultation, learn more about our extensive product line that can help you with protein expression services and purification, and explore fresh automation solutions to fulfill all your lab requirements.
