Epitope discovery for a synthetic polymer nanoparticle: A new strategy for developing a peptide tag
| Autor: | Linda F. Epstein, Keiichi Yoshimatsu, Yu Hoshino, John M. Beierle, Paul E. Rose, Kenneth J. Shea, Les P. Miranda, Philip Tagari, Tomohiko Yamazaki, Yusuke Yonamine |
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| Jazyk: | angličtina |
| Rok vydání: | 2014 |
| Předmět: |
Strep-tag
Polymers Surface Properties Peptide Bioengineering 02 engineering and technology Protein tag 010402 general chemistry 01 natural sciences Biochemistry Catalysis Epitopes Colloid and Surface Chemistry FLAG-tag Nanotechnology Particle Size chemistry.chemical_classification biology Molecular Structure Chemistry Communication General Chemistry 021001 nanoscience & nanotechnology Avidin Combinatorial chemistry Isotope-coded affinity tag Recombinant Proteins 0104 chemical sciences Biotinylation Chemical Sciences biology.protein Nanoparticles 0210 nano-technology Peptides Myc-tag |
| Zdroj: | Yoshimatsu, K; Yamazaki, T; Hoshino, Y; Rose, PE; Epstein, LF; Miranda, LP; et al.(2014). Epitope discovery for a synthetic polymer nanoparticle: A new strategy for developing a peptide tag. Journal of the American Chemical Society, 136(4), 1194-1197. doi: 10.1021/ja410817p. UC Irvine: Retrieved from: http://www.escholarship.org/uc/item/9v59x55n Journal of the American Chemical Society Journal of the American Chemical Society, vol 136, iss 4 |
| DOI: | 10.1021/ja410817p. |
| Popis: | We describe a novel epitope discovery strategy for creating an affinity agent/peptide tag pair. A synthetic polymer nanoparticle (NP) was used as the "bait" to catch an affinity peptide tag. Biotinylated peptide tag candidates of varied sequence and length were attached to an avidin platform and screened for affinity against the polymer NP. NP affinity for the avidin/peptide tag complexes was used to provide insight into factors that contribute NP/tag binding. The identified epitope sequence with an optimized length (tMel-tag) was fused to two recombinant proteins. The tagged proteins exhibited higher NP affinity than proteins without tags. The results establish that a fusion peptide tag consisting of optimized 15 amino acid residues can provide strong affinity to an abiotic polymer NP. The affinity and selectivity of NP/tMel-tag interactions were exploited for protein purification in conjunction with immobilized metal ion/His6-tag interactions to prepare highly purified recombinant proteins. This strategy makes available inexpensive, abiotic synthetic polymers as affinity agents for peptide tags and provides alternatives for important applications where more costly affinity agents are used. © 2014 American Chemical Society. |
| Databáze: | OpenAIRE |
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