Recombinant Protein Purification
Recombinant Protein Purification - Related Videos
07:44
High-throughput Purification of Affinity-tagged Recombinant Proteins
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Cited by 1 •
2012
We describe a method for the affinity-tagged purification of recombinant proteins using liquid-handling robotics. This method is generally applicable to the small-scale purification of soluble His-tagged proteins in a high-throughput format.
02:01
Purification of Bacteria-Derived Recombinant P Domain Proteins of Human Norovirus
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2025
Source: Leuthold, M. M., et al. Production of Human Norovirus Protruding Domains in E. coli for X-ray Crystallography. J. Vis. Exp. (2016).This video demonstrates the purification of recombinant human norovirus P domain using size exclusion chromatography, highlighting the separation of the target protein from higher and lower molecular weight impurities based on differential pore accessibility. The process is monitored by UV absorbance and confirmed through SDS-PAGE analysis of eluted...
02:54
An Affinity Chromatography Technique for the Purification of a Recombinant Bacterial Protein
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2025
Source: Becker, B., et al. A Simple Fluorescence-based Reporter Assay to Identify Cellular Components Required for Ricin Toxin A Chain (RTA) Trafficking in Yeast. J. Vis. Exp. (2017).This video demonstrates the purification of a polyhistidine-tagged recombinant protein using nickel-based affinity chromatography. It outlines key steps, including bacterial lysis by sonication, clarification and filtration of the lysate, and selective binding and elution of the target protein using imidazole.
03:59
Isolation and Purification of Recombinant Myelin Oligodendrocyte Glycoproteins
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2025
Source: Jain, R. W., et al. Simple and Efficient Production and Purification of Mouse Myelin Oligodendrocyte Glycoprotein for Experimental Autoimmune Encephalomyelitis Studies. J. Vis. Exp. (2016)This video demonstrates the isolation and purification of MOGtag, a recombinant Myelin Oligodendrocyte Glycoprotein (MOG) used in Experimental Autoimmune Encephalomyelitis (EAE) studies. The protocol involves bacterial lysis, sonication, centrifugation, and nickel affinity chromatography to obtain...
07:35
Non-chromatographic Purification of Recombinant Elastin-like Polypeptides and their Fusions with Peptides and Proteins from Escherichia coli
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Cited by 48 •
2014
Elastin-like polypeptides are stimulus-responsive biopolymers with applications ranging from recombinant protein purification to drug delivery. This protocol describes the purification and characterization of elastin-like polypeptides and their peptide or protein fusions from Escherichia coli using their lower critical solution temperature phase transition behavior as a simple alternative to chromatography.
FAQs
Affinity chromatography uses a selective interaction between a tagged protein and a ligand attached to a resin. When the prepared protein mixture contacts the resin, the tagged target binds while other material can be separated. This selectivity helps enrich the intended protein for later structural, functional, immunological, or infection-related studies.
The bound protein is released by changing buffer conditions or adding a competing molecule. Either approach disrupts the interaction between the tag and the resin-associated ligand, allowing the target protein to be collected. The choice of release condition matters because it determines whether the protein can be recovered for subsequent assays or applications.
Cell lysis makes the recombinant protein accessible by breaking open the genetically engineered cells that produce it. The resulting cellular material can then be processed with a separation method such as affinity chromatography. Effective preparation is therefore an important early step in obtaining protein suitable for studying structure, function, immune recognition, or pathogen-related biology.
A typical workflow begins with genetically engineered cells, followed by cell lysis to release their protein contents. The preparation is then subjected to a separation step, commonly affinity chromatography, where the tagged target binds a resin ligand. Finally, changing buffer conditions or adding a competing molecule releases the protein for collection and analysis.
Purified proteins can serve as antigens, antibodies, enzymes, diagnostic reagents, or vaccine components. Their defined composition supports experiments focused on immune recognition, pathogen biology, host-pathogen interactions, and therapeutic responses. The appropriate use depends on the protein's role in the planned assay or research question.
Protein quality and purity influence the reliability of assays that examine immune recognition, pathogen biology, host-pathogen interactions, or therapeutic responses. A preparation that is sufficiently purified provides a more dependable research reagent, whereas inconsistent quality can make experimental observations harder to interpret. Purification is therefore closely linked to confidence in study results.