Recombinant proteins are proteins that are produced by genetically engineered cells, which have been modified to contain a foreign gene encoding the protein of interest. These proteins are often used for research, diagnostic, and therapeutic purposes. The production of recombinant proteins involves introducing a gene encoding the desired protein into a suitable host organism, such as bacteria, yeast, or mammalian cells, which then express the protein using their own cellular machinery.
The main steps in the production of recombinant proteins are:
- Gene cloning: The gene encoding the protein of interest is isolated from its source organism or synthesized de novo. This gene is then inserted into a suitable expression vector, which is a piece of DNA designed to carry and promote the expression of the target gene.
- Transformation or transfection: The expression vector containing the target gene is introduced into a compatible host organism, such as Escherichia coli (E. coli) bacteria, yeast cells, or mammalian cells.
- Expression: The host organism’s cellular machinery is used to transcribe and translate the foreign gene, producing the desired protein. Various factors, including the choice of host organism, promoter, and codon optimization, can influence the efficiency of protein expression.
- Protein purification: The recombinant protein is extracted from the host organism and purified using a series of chromatographic and other separation techniques. The purification process aims to remove contaminants, such as host proteins, DNA, and endotoxins, to obtain a highly purified product suitable for downstream applications.
Recombinant protein technology has revolutionized the fields of biotechnology and medicine, enabling the large-scale production of proteins for research and therapeutic use. Examples of recombinant proteins used as therapeutics include insulin, erythropoietin, and monoclonal antibodies.
