Fluorescent-conjugated polymer superquenching facilitates highly sensitive detection of proteases

被引：199

作者：

Kumaraswamy, S ^{[1
]}

Bergstedt, T ^{[1
]}

Shi, XB ^{[1
]}

Rininsland, F ^{[1
]}

Kushon, S ^{[1
]}

Xia, WS ^{[1
]}

Ley, K ^{[1
]}

Achyuthan, K ^{[1
]}

McBranch, D ^{[1
]}

Whitten, D ^{[1
]}

机构：

[1] QTL Biosyst, Santa Fe, NM 87507 USA

来源：

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA | 2004年 / 101卷 / 20期

关键词：

D O I：

10.1073/pnas.0402367101

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

Sensor formats have been developed for detecting the activity of proteolytic enzymes based on fluorescent conjugated polymer superquenching. These sensors employ a reactive peptide sequence within a tether linking a quencher to a biotin. The peptide binds to sensors containing colocated biotin-binding protein and fluorescent polymer by means of biotin-biotin binding protein interactions, resulting in a strong quenching of polymer fluorescence, Enzyme-mediated cleavage of the peptide results in a reversal of the fluorescence quenching. These assays for protease activity are simple, sensitive, fast, and have the specificity required for screening chemical libraries for novel protease inhibitors in a high-throughput screening assay environment. These assays have been demonstrated for enterokinase, caspase-3/7, and beta-secretase.

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页码：7511 / 7515

页数：5

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