共 31 条
RAMPED-UP NMR: Multiplexed NMR-based screening for drug discovery
被引:40
作者:

Zartler, ER
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机构: Eli Lilly & Co, Lilly Corp Ctr, Lilly Res Labs,Biol Res Technol & Prot, Discovery Chem Res & Technol, Indianapolis, IN 46285 USA

Hanson, J
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机构: Eli Lilly & Co, Lilly Corp Ctr, Lilly Res Labs,Biol Res Technol & Prot, Discovery Chem Res & Technol, Indianapolis, IN 46285 USA

Jones, BE
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机构: Eli Lilly & Co, Lilly Corp Ctr, Lilly Res Labs,Biol Res Technol & Prot, Discovery Chem Res & Technol, Indianapolis, IN 46285 USA

Kline, AD
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机构: Eli Lilly & Co, Lilly Corp Ctr, Lilly Res Labs,Biol Res Technol & Prot, Discovery Chem Res & Technol, Indianapolis, IN 46285 USA

Martin, G
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机构: Eli Lilly & Co, Lilly Corp Ctr, Lilly Res Labs,Biol Res Technol & Prot, Discovery Chem Res & Technol, Indianapolis, IN 46285 USA

Mo, HP
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机构: Eli Lilly & Co, Lilly Corp Ctr, Lilly Res Labs,Biol Res Technol & Prot, Discovery Chem Res & Technol, Indianapolis, IN 46285 USA

Shapiro, MJ
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机构:
Eli Lilly & Co, Lilly Corp Ctr, Lilly Res Labs,Biol Res Technol & Prot, Discovery Chem Res & Technol, Indianapolis, IN 46285 USA Eli Lilly & Co, Lilly Corp Ctr, Lilly Res Labs,Biol Res Technol & Prot, Discovery Chem Res & Technol, Indianapolis, IN 46285 USA

Wang, R
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机构: Eli Lilly & Co, Lilly Corp Ctr, Lilly Res Labs,Biol Res Technol & Prot, Discovery Chem Res & Technol, Indianapolis, IN 46285 USA

Wu, HP
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机构: Eli Lilly & Co, Lilly Corp Ctr, Lilly Res Labs,Biol Res Technol & Prot, Discovery Chem Res & Technol, Indianapolis, IN 46285 USA

Yan, JL
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机构: Eli Lilly & Co, Lilly Corp Ctr, Lilly Res Labs,Biol Res Technol & Prot, Discovery Chem Res & Technol, Indianapolis, IN 46285 USA
机构:
[1] Eli Lilly & Co, Lilly Corp Ctr, Lilly Res Labs,Biol Res Technol & Prot, Discovery Chem Res & Technol, Indianapolis, IN 46285 USA
[2] Roche Diagnost Corp, Roche Prot Express Grp, Indianapolis, IN 46250 USA
关键词:
D O I:
10.1021/ja0348593
中图分类号:
O6 [化学];
学科分类号:
0703 ;
摘要:
The crucial step in drug discovery is the identification of a lead compound from a vast chemical library by any number of screening techniques. NMR-based screening has the advantage of directly detecting binding of a compound to the target. The spectra resulting from these screens can also be very complex and difficult to analyze, making this an inefficient process. We present here a method, RAMPED-UP NMR, (Rapid Analysis and Multiplexing of Experimentally Discriminated Uniquely Labeled Proteins using NMR) which generates simple spectra which are easy to interpret and allows several proteins to be screened simultaneously. In this method, the proteins to be screened are uniquely labeled with one amino acid type. There are several benefits derived from this unique labeling strategy: the spectra are greatly simplified, resonances that are most likely to be affected by binding are the only ones observed, and peaks that yield little or no information upon binding are eliminated, allowing the analysis of multiple proteins easily and simultaneously. We demonstrate the ability of three different proteins to be analyzed simultaneously for binding to two different ligands. This method will have significant impact in the use of NMR spectroscopy for both the lead generation and lead optimization phases of drug discovery by its ability to increase screening throughput and the ability to examine selectivity. To the best of our knowledge, this is the first time in any format that multiple proteins can be screened in one tube.
引用
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页码:10941 / 10946
页数:6
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机构: Novo Nordisk AS, Prot Chem, DK-2880 Bagsvaerd, Denmark

Ge, Y
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机构: Novo Nordisk AS, Prot Chem, DK-2880 Bagsvaerd, Denmark

Holsworth, DD
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机构: Novo Nordisk AS, Prot Chem, DK-2880 Bagsvaerd, Denmark

Newman, NJ
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机构: Novo Nordisk AS, Prot Chem, DK-2880 Bagsvaerd, Denmark

Moller, NPH
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机构: Novo Nordisk AS, Prot Chem, DK-2880 Bagsvaerd, Denmark