Recombinant protein production uses DNA techniques to produce proteins in large quantities for laboratory and industrial use. It is used to produce human growth hormone and insulin, and involves inserting genes into vectors and inducing overexpression in host cells. Expression vectors have promoters to make large amounts of protein, and purification is facilitated with tags that bind to matrices. Mammalian proteins often require modification and are produced in insect or mammalian cells. Biotechnology companies specialize in this production.
Recombinant protein production is the expression of proteins that have been produced by recombinant DNA techniques. This process makes it possible to produce these substances in large quantities. Such mass production is carried out both for laboratory study and for industrial production.
This technique is often used to produce human growth hormone and insulin. Obtaining human growth hormone through the production of recombinant proteins is a huge improvement over obtaining it from cadavers because the presence of cadaveric-obtained proteins occasionally resulted in disease transmission. Making insulin this way is also beneficial because it made it possible to produce insulin variants that have different pharmacological actions in the body.
Proteins are chains of amino acids, encoded by DNA. The genes that code for these proteins are inserted into special vectors, or units of DNA. Carriers are chosen that will produce large quantities of the desired protein. This is known as overexpression.
Overexpression occurs in special host cells. Sometimes the hosts are bacteria or yeast. In cases where the proteins originate from mammals, the hosts are often insect or mammalian cell lines. A number of kits are commercially available to facilitate both the cloning of the gene and the subsequent production of recombinant proteins.
These kits have special vectors called expression vectors which have a special promoter to make large amounts of protein. A promoter is the section of DNA that drives the production of the gene sequence that follows it. Often, these expression vectors can be turned off and are inducible. Especially with bacterial hosts, making too much protein at once can be toxic, inhibiting the growth of bacteria.
There are several ways to induce expression. In both, the bacteria grew to a certain density. Then a compound is added for induction or the temperature is shifted to that where the promoter is active.
To facilitate the purification of proteins from bacteria, cloning is often done so that there is a tag on the protein that binds to a matrix. This separates the protein from the cellular debris. For example, a tag of histidine molecules on the protein will bind to a nickel column. Once the protein is bound, the tag is detached, leaving pure protein which can be eluted from the column. It can take years to purify a protein using traditional methods.
An additional factor to consider is whether the protein requires modification after its initial production. This is often the case with mammalian proteins. Bacteria often do not modify such proteins adequately, so overexpression of these more advanced proteins is often carried out in insect or mammalian cells. A number of biotechnology companies specialize in the production of recombinant proteins.
Protect your devices with Threat Protection by NordVPN
