Abstract
Applied in tandem, elastin-like polypeptides (ELPs) and the sortase A (SrtA) transpeptidase from Staphylococcus aureus provide a general method for chromatography-free purification of tag-free recombinant proteins and optional, site-specific and homogeneous conjugation of the protein to a small molecule. This system provides an efficient, practical mechanism for generating bioactive proteins and protein-small-molecule combination therapeutics at high yields and purities.
Publication types
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Research Support, N.I.H., Extramural
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Research Support, Non-U.S. Gov't
MeSH terms
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Aminoacyltransferases / genetics
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Aminoacyltransferases / metabolism*
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Animals
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Antineoplastic Agents / chemistry
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Antineoplastic Agents / metabolism
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Antineoplastic Agents / toxicity
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Bacterial Proteins / genetics
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Bacterial Proteins / metabolism*
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Cell Line, Tumor
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Cell Survival / drug effects
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Cysteine Endopeptidases / genetics
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Cysteine Endopeptidases / metabolism*
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Elastin / genetics
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Elastin / metabolism*
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Escherichia coli / metabolism
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Humans
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Mice
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Mutagenesis, Site-Directed
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Recombinant Fusion Proteins / genetics
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Recombinant Fusion Proteins / metabolism*
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Staphylococcus aureus / enzymology
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TNF-Related Apoptosis-Inducing Ligand / genetics
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TNF-Related Apoptosis-Inducing Ligand / metabolism
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Tumor Necrosis Factor-alpha / genetics
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Tumor Necrosis Factor-alpha / metabolism
Substances
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Antineoplastic Agents
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Bacterial Proteins
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Recombinant Fusion Proteins
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TNF-Related Apoptosis-Inducing Ligand
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Tumor Necrosis Factor-alpha
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Elastin
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Aminoacyltransferases
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sortase A
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Cysteine Endopeptidases