Multiple ligand detection and affinity measurement by ultrafiltration and mass spectrometry analysis applied to fragment mixture screening

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	Multiple ligand detection and affinity measurement by ultrafiltration and mass spectrometry analysis applied to fragment mixture screening
	Qin, Shanshan 1,2; Ren, Yiran 2; Fu, Xu 2; Shen, Jie 1; Chen, Xin 2; Wang, Quan 2; Bi, Xin 3; Liu, Wenjing 2; Li, Lixin 2; Liang, Guangxin 3
刊名	ANALYTICA CHIMICA ACTA
	2015-07-30
卷号	886 页码:98-106
关键词	Ultrafiltration-LC/MS Multiple ligand detection Affinity measurement Fragment library screening NS5B
英文摘要	Binding affinity of a small molecule drug candidate to a therapeutically relevant biomolecular target is regarded the first determinant of the candidate's efficacy. Although the ultrafiltration-LC/MS (UF-LC/MS) assay enables efficient ligand discovery for a specific target from a mixed pool of compounds, most previous analysis allowed for relative affinity ranking of different ligands. Moreover, the reliability of affinity measurement for multiple ligands with UF-LC/MS has hardly been strictly evaluated. In this study, we examined the accuracy of K-d determination through UF-LC/MS by comparison with classical ITC measurement. A single-point Kd calculation method was found to be suitable for affinity measurement of multiple ligands bound to the same target when binding competition is minimized. A second workflow based on analysis of the unbound fraction of compounds was then developed, which simplified sample preparation as well as warranted reliable ligand discovery. The new workflow implemented in a fragment mixture screen afforded rapid and sensitive detection of low-affinity ligands selectively bound to the RNA polymerase NS5B of hepatitis C virus. More importantly, ligand identification and affinity measurement for mixture-based fragment screens by UF-LC/MS were in good accordance with single ligand evaluation by conventional SPR analysis. This new approach is expected to become a valuable addition to the arsenal of high-throughput screening techniques for fragment-based drug discovery. (C) 2015 Elsevier B.V. All rights reserved.
WOS标题词	Science & Technology ; Physical Sciences
类目[WOS]	Chemistry, Analytical
研究领域[WOS]	Chemistry
关键词[WOS]	COLLISION-INDUCED DISSOCIATION ; CARBONIC-ANHYDRASE-I ; CHROMATOGRAPHY/MASS SPECTROMETRY ; DRUG DISCOVERY ; LEAD DISCOVERY ; BINDING AFFINITIES ; COMPOUND MIXTURES ; ELECTRON-TRANSFER ; RNA-POLYMERASE ; INHIBITORS
收录类别	SCI
语种	英语
WOS记录号	WOS:000360643800011
内容类型	期刊论文
源URL	[http://124.16.173.210/handle/834782/1518]
专题	天津工业生物技术研究所_生物质谱与定量生物学研究水雯箐_期刊论文
作者单位	1.Chinese Acad Sci, Tianjin Inst Ind Biotechnol, Key Lab Syst Microbial Biotechnol, Tianjin 300308, Peoples R China 2.Tianjin Joint Acad Biotechnol & Med, High Throughput Mol Drug Discovery Ctr, Tianjin 300457, Peoples R China 3.Nankai Univ, Inst Elementoorgan Chem, Tianjin 300071, Peoples R China
推荐引用方式 GB/T 7714	Qin, Shanshan,Ren, Yiran,Fu, Xu,et al. Multiple ligand detection and affinity measurement by ultrafiltration and mass spectrometry analysis applied to fragment mixture screening[J]. ANALYTICA CHIMICA ACTA,2015,886:98-106.
APA	Qin, Shanshan.,Ren, Yiran.,Fu, Xu.,Shen, Jie.,Chen, Xin.,...&Shui, Wenqing.(2015).Multiple ligand detection and affinity measurement by ultrafiltration and mass spectrometry analysis applied to fragment mixture screening.ANALYTICA CHIMICA ACTA,886,98-106.
MLA	Qin, Shanshan,et al."Multiple ligand detection and affinity measurement by ultrafiltration and mass spectrometry analysis applied to fragment mixture screening".ANALYTICA CHIMICA ACTA 886(2015):98-106.