Recombinant Proteins: Engineered Biomolecules Powering Research and Therapeutics - Healthcare-netizens/arpita-kamat GitHub Wiki
Recombinant proteins are proteins that are produced using recombinant DNA technology. This involves manipulating the genetic material of an organism to introduce and express a gene encoding the desired protein in a host cell. The host cell, which can be bacteria, yeast, insect cells, or mammalian cells, then acts as a biological factory, producing large quantities of the recombinant protein. This technology has revolutionized various fields, providing a reliable and scalable way to produce proteins that may be scarce or difficult to obtain naturally.
The process of producing a recombinant protein typically involves several key steps:
Gene Cloning: The gene encoding the target protein is isolated from its source organism (e.g., human, animal, plant) and inserted into a vector, such as a plasmid or a virus. The vector acts as a carrier to deliver the gene into the host cell.
Transformation or Transfection: The recombinant vector containing the gene of interest is introduced into the host cells. This process is called transformation in bacteria and yeast, and transfection in insect and mammalian cells.
Selection of Transformed/Transfected Cells: Only the host cells that have successfully taken up the recombinant vector and integrated the gene into their genome (or maintain it as an extrachromosomal element) are selected. This is often achieved using selectable markers present on the vector, such as antibiotic resistance genes.
Protein Expression: The selected host cells are cultured under conditions that allow for the expression of the recombinant gene. This involves providing the necessary nutrients, temperature, and sometimes specific inducing agents that trigger the transcription and translation of the target protein.
Protein Purification: Once the recombinant protein is produced by the host cells, it needs to be isolated and purified from the complex mixture of cellular components. This often involves a series of chromatographic techniques, such as affinity chromatography (using a specific binding partner for the protein), ion exchange chromatography (separating proteins based on charge), and size exclusion chromatography (separating proteins based on size).
Characterization and Quality Control: The purified recombinant protein undergoes rigorous characterization to confirm its identity, purity, activity, and stability. Various analytical techniques, such as SDS-PAGE, Western blotting, mass spectrometry, and functional assays, are employed for this purpose.
Recombinant protein technology offers numerous advantages over traditional methods of protein production. It allows for the production of large quantities of virtually any protein, even those that are naturally rare or difficult to isolate. It enables the engineering of proteins with modified properties, such as increased stability, altered activity, or the addition of tags for purification or detection. Furthermore, it provides a more controlled and consistent production process, which is particularly important for therapeutic applications.
The applications of recombinant proteins are vast and continue to expand, impacting fields ranging from basic research to medicine and industry.
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