Abstract
The affinity tags are unique proteins/peptides that are attached at the N- or C-terminus of the recombinant proteins. These tags help in protein purification. Additionally, some affinity tags also serve a dual purpose as solubility enhancers for challenging protein targets. By applying a combinatorial approach, carefully chosen affinity tags designed in tandem have proven to be very successful in the purification of single proteins or multi-protein complexes. In this mini-review, the key features of the most commonly used affinity tags are discussed. The affinity tags have been classified into two significant categories, epitope tags, and protein/domain tags. The epitope tags are generally small peptides with high affinity towards a chromatography resin. The protein/domain tags often perform double duty as solubility enhancers as well as aid in affinity purification. Finally, protease-based affinity tag removal strategies after purification are discussed.
Keywords:
Proteins; affinity tag; chromatography; protease; purification; solubility.
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MeSH terms
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Chromatography, Affinity / methods*
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Epitopes / chemistry
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Epitopes / metabolism
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Escherichia coli / genetics*
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Escherichia coli / metabolism
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Escherichia coli Proteins / genetics
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Escherichia coli Proteins / metabolism
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Gene Expression
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Glutathione Transferase / genetics
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Glutathione Transferase / metabolism
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Hemagglutinin Glycoproteins, Influenza Virus / genetics
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Hemagglutinin Glycoproteins, Influenza Virus / metabolism
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Histidine / genetics*
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Histidine / metabolism
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Maltose-Binding Proteins / genetics
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Maltose-Binding Proteins / metabolism
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Oligopeptides / genetics*
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Oligopeptides / metabolism
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Peptide Hydrolases / chemistry
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Peptides / chemistry
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Peptides / genetics
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Peptides / metabolism
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Protein Domains
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Proteolysis
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Recombinant Fusion Proteins / chemistry*
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Recombinant Fusion Proteins / genetics
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Recombinant Fusion Proteins / isolation & purification
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Recombinant Fusion Proteins / metabolism
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SUMO-1 Protein / genetics
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SUMO-1 Protein / metabolism
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Solubility
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Staining and Labeling / methods*
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Transcriptional Elongation Factors / genetics
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Transcriptional Elongation Factors / metabolism
Substances
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Epitopes
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Escherichia coli Proteins
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Hemagglutinin Glycoproteins, Influenza Virus
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His-His-His-His-His-His
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Maltose-Binding Proteins
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Oligopeptides
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Peptides
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Recombinant Fusion Proteins
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SUMO-1 Protein
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Transcriptional Elongation Factors
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nusA protein, E coli
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Histidine
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Glutathione Transferase
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Peptide Hydrolases