Epitope discovery for a synthetic polymer nanoparticle: a new strategy for developing a peptide tag.
Epitope discovery for a synthetic polymer nanoparticle: a new strategy for developing a peptide tag.
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DOI：
10.1021/ja410817p
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发表时间：
2014-01-29
影响因子：
15
通讯作者：
Shea KJ
中科院分区：
文献类型：
--
作者：
Yoshimatsu K;Yamazaki T;Hoshino Y;Rose PE;Epstein LF;Miranda LP;Tagari P;Beierle JM;Yonamine Y;Shea KJ
我们描述了一种新的表位发现策略，用于创建亲和剂/肽标签对。合成聚合物纳米颗粒（NP）作为“诱饵”捕获亲和肽标签。将不同序列和长度的候选生物素化肽标签附着在亲和素平台上，并筛选其对聚合物NP的亲和力。利用亲和素/肽标签复合物的NP亲和力来深入了解促成NP/标签结合的因素。将确定的表位序列与优化长度（tMel-tag）融合到两个重组蛋白上。标记的蛋白比未标记的蛋白表现出更高的NP亲和力。结果表明，由优化后的15个氨基酸残基组成的融合肽标签对非生物聚合物NP具有较强的亲和力。利用NP/tMel-tag相互作用的亲和性和选择性，结合固定化金属离子/His6-tag相互作用，制备高纯度的重组蛋白。这一策略使得廉价的非生物合成聚合物成为肽标签的亲和剂，并为使用更昂贵的亲和剂的重要应用提供了替代方案。
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.
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影响因子：
7.8
作者：
Rana, Subinoy;Yeh, Yi-Cheun;Rotello, Vincent M.
通讯作者：
Rotello, Vincent M.
影响因子：
18.3
作者：
Kim, Sung Tae;Saha, Krishnendu;Kim, Chaekyu;Rotello, Vincent M.
通讯作者：
Rotello, Vincent M.
影响因子：
15
作者：
Fischer, NO;Verma, A;Rotello, VM
通讯作者：
Rotello, VM
DOI：
10.1038/nbt1088-1204
发表时间：
1988-10-01
期刊：
BIO-TECHNOLOGY
影响因子：
--
作者：
HOPP, TP;PRICKETT, KS;CONLON, PJ
通讯作者：
CONLON, PJ
影响因子：
3.9
作者：
Ito, S;Ogawa, K;Kokufuta, E
通讯作者：
Kokufuta, E