Applications of novel affinity cassette methods: use of peptide fusion handles for the purification of recombinant proteins

Hearn, Milton Thomas William; Acosta, Danilo B

doi:10.1002/jmr.555

Cited by 101 publications

(84 citation statements)

References 316 publications

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“…Detailed descriptions of these affinity tags can be found elsewhere in comprehensive reviews [364][365][366][367][368]. Recently, affinity tags that can specifically and strongly bind to non-biological molecules, including bulk materials, have been developed by phage display and other combinatorial techniques.…”

Section: Micro-and Nano-structures-mentioning

confidence: 99%

Peptide-based biopolymers in biomedicine and biotechnology

Chow

Nunalee

Lim

et al. 2008

Materials Science and Engineering: R: Reports

280

231

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Peptides are emerging as a new class of biomaterials due to their unique chemical, physical, and biological properties. The development of peptide-based biomaterials is driven by the convergence of protein engineering and macromolecular self-assembly. This review covers the basic principles, applications, and prospects of peptide-based biomaterials. We focus on both chemically synthesized and genetically encoded peptides, including poly-amino acids, elastin-like polypeptides, silk-like polymers and other biopolymers based on repetitive peptide motifs. Applications of these engineered biomolecules in protein purification, controlled drug delivery, tissue engineering, and biosurface engineering are discussed.

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Section: Micro-and Nano-structures-mentioning

confidence: 99%

Peptide-based biopolymers in biomedicine and biotechnology

Chow

Nunalee

Lim

et al. 2008

Materials Science and Engineering: R: Reports

280

231

View full text Add to dashboard Cite

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“…Comprehensive reviews of affinity tags have been published recently [17,18]. Although many different tags have been described in the literature, few of them have been tested in a high-throughput context.…”

Section: Enhancing the Solubility Of Recombinant Proteinsmentioning

confidence: 99%

Making the most of affinity tags

Waugh

2005

Trends in Biotechnology

478

324

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“…These tags are widely used for protein localization or to enhance the solubility of the target protein (Huh et al, 2003;Smyth et al, 2003). Affinity tags have been fused to either the N-or C-terminus of target proteins (Hearn and Acosta, 2001;Arnau et al, 2006) to insure the likelihood of success in protein detection and purification.…”

Section: Introductionmentioning

confidence: 99%

The utility of affinity-tags for detection of a streptococcal protein from a variety of streptococcal species

Zhou

Fives-Taylor

2008

Journal of Microbiological Methods

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There is no systematic examination of affinity tag utility in Gram-positive bacteria, which limits the investigation of protein function in this important group of bacteria as specific antibodies for many of native proteins are generally not available. In this study, we utilized an E. coli-streptococcal shuttle vector pVT1666 and constructed two sets of expression plasmids pVPT-CTag and pVPT-NTag, with each set containing five affinity tags (GST, GFP, HSV, T7 and Nano) that can be fused to either the C-or N-terminus of a target protein. A putative glycosyltransferase (Gtf2) essential for Fap1 glycosylation was used to demonstrate the utility of the cassettes in detection of Gtf2 fusion proteins, and the biological relevance of the proteins in our working strain Streptococcus parasanguinis. GFP and T7 tags were readily expressed in S. parasanguinis as either an N-or C-terminal fusion to Gtf2. Only the C-terminal fusion of GST and HSV were able to be identified in S. parasanguinis. The Nano tag was not detected in either E. coli or S. parasanguinis. Genetic complementation experiments indicated that all the tagged Gtf2 fusion proteins could restore the Gtf2 function in the null mutant except for the Nano-tagged Gtf2 at its N-terminal fusion. Using a T7-tagged Gtf2 fusion construct, we demonstrated that the fusion cassette is also useful in detection of the fusion tag expression in other streptococci including S. mutans, S. pneumoniae and S. sanguinis. Therefore, the expression cassettes we constructed will be a useful tool not only to investigate protein-protein interactions in Fap1 biogenesis in S. parasanguinis, but also to study protein functions in other gram-positive bacteria in which pVT1666 replicates.

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Applications of novel affinity cassette methods: use of peptide fusion handles for the purification of recombinant proteins

Cited by 101 publications

References 316 publications

Peptide-based biopolymers in biomedicine and biotechnology

Peptide-based biopolymers in biomedicine and biotechnology

Making the most of affinity tags

The utility of affinity-tags for detection of a streptococcal protein from a variety of streptococcal species

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