Supra-blot: an accurate and reliable assay for detecting target proteins with a synthetic host molecule-enzyme hybrid.,Chemical Communications

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Supra-blot: an accurate and reliable assay for detecting target proteins with a synthetic host molecule-enzyme hybrid.
Chemical Communications ( IF 4.3 ) Pub Date : 2020-01-13 , DOI: 10.1039/c9cc09699j
Gihyun Sung ₁ , Song-Yi Lee ₂ , Myeong-Gyun Kang ₃ , Kyung Lock Kim ₄ , Jaeyeon An ₅ , Jaehwan Sim ₆ , Sungwan Kim ₇ , Seungjoon Kim ₈ , Jaewon Ko ₈ , Hyun-Woo Rhee ₂ , Kyeng Min Park ₄ , Kimoon Kim ₉

Affiliation

Center for Self-assembly and Complexity, Institute for Basic Science (IBS), Pohang, 37673, Republic of Korea. and Division of Advanced Materials Science (AMS), Pohang University of Science and Technology (POSTECH), Pohang, 37673, Republic of Korea.
Department of Chemistry, Seoul National University, Seoul, 08826, Republic of Korea.
Department of Chemistry, Ulsan National Institute of Science and Technology (UNIST), Ulsan, 44919, Republic of Korea.
Center for Self-assembly and Complexity, Institute for Basic Science (IBS), Pohang, 37673, Republic of Korea.
Department of Chemistry, Pohang University of Science and Technology (POSTECH), Pohang, Korea.
School of Interdisciplinary Bioscience and Bioengineering (i-bio), Pohang University of Science and Technology (POSTECH), Pohang, Korea.
Center for Self-assembly and Complexity, Institute for Basic Science (IBS), Pohang, 37673, Republic of Korea. and Department of Chemistry, Pohang University of Science and Technology (POSTECH), Pohang, Korea.
Department of Brain and Cognitive Sciences, Daegu Gyeongbuk Institute of Science and Technology (DGIST), Daegu 42988, Republic of Korea.
Center for Self-assembly and Complexity, Institute for Basic Science (IBS), Pohang, 37673, Republic of Korea. and Division of Advanced Materials Science (AMS), Pohang University of Science and Technology (POSTECH), Pohang, 37673, Republic of Korea. and Department of Chemistry, Pohang University of Science and Technology (POSTECH), Pohang, Korea and School of Interdisciplinary Bioscience and Bioengineering (i-bio), Pohang University of Science and Technology (POSTECH), Pohang, Korea.

In accordance with the rapid increase in demand for selective and spatial chemical tagging, and accurate detection of proteins of interest, we develop a sensitive protein detection method, termed "Supra-blot" capitalizing on high-affinity host-guest interaction between cucurbit[7]uril (CB[7]) and adamantylammonium (AdA). The method can directly detect chemically tagged proteins without false-positive signals caused by endogenous biomolecules. Not only a single specific protein, but also spatially localized proteins in cells were labeled with AdA, and selectively detected by a host molecule-enzyme hybrid, CB[7]-conjugated horseradish peroxidase (CB[7]-HRP) generating amplified chemiluminescence signals. This study shows the great potential of Supra-blot for accurate and reliable detection of proteins of interest in cells.

中文翻译：

超印迹：使用合成的宿主分子-酶杂交体检测靶蛋白的准确可靠的测定法。

为了满足对选择性和空间化学标签的需求的快速增长以及对目标蛋白质的准确检测的需求，我们开发了一种敏感的蛋白质检测方法，称为“超印迹”，利用葫芦丝之间的高亲和性宿主-客体相互作用[7] ] uril（CB [7]）和金刚烷铵（AdA）。该方法可以直接检测化学标记的蛋白质，而没有由内源性生物分子引起的假阳性信号。用AdA不仅标记单个特异性蛋白，还标记细胞中空间定位的蛋白，并通过宿主分子-酶杂交体CB [7]-缀合的辣根过氧化物酶（CB [7] -HRP）选择性检测，产生放大的化学发光信号。这项研究显示了Supra印迹在准确可靠地检测细胞中目标蛋白质方面的巨大潜力。

更新日期：2020-01-13

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