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

Analytica Chimica Acta
Shanshan QinWenqing Shui

Abstract

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 K(d) 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 mixtu...Continue Reading

References

Dec 26, 2001·Rapid Communications in Mass Spectrometry : RCM·Paul A Wabnitz, Joseph A Loo
Apr 5, 2002·Journal of Medicinal Chemistry·Susan L McGovernBrian K Shoichet
Sep 29, 2004·Journal of Biomolecular Screening·Hartmut ZehenderLorenz M Mayr
Oct 12, 2004·Journal of Biomolecular Screening·Anthony M SmithShu-Gui Huang
Nov 25, 2004·Antimicrobial Agents and Chemotherapy·Anita Y M HoweJohn F O'Connell
Jul 19, 2005·Drug Discovery Today·Robin A E CarrDavid C Rees
Nov 1, 2005·Analytical Chemistry·Xun Cheng, Richard B van Breemen
Aug 4, 2006·Journal of Medicinal Chemistry·Alessio CiulliChris Abell
Feb 10, 2007·Nature Reviews. Drug Discovery·Philip J Hajduk, Jonathan Greer
Apr 5, 2007·Journal of the American Chemical Society·D K SrivastavaDavid W Christianson
May 16, 2007·Analytical Chemistry·D Allen AnnisWilliam T Windsor
Oct 13, 2007·Current Opinion in Chemical Biology·D Allen AnnisCharles E Whitehurst
Nov 14, 2007·Analytical Chemistry·Dongting LiuRichard B van Breemen
Apr 11, 2008·Bioorganic & Medicinal Chemistry Letters·Stephen S AntonysamyTobi Wright
Jun 25, 2008·Annual Review of Biophysics·Jonathan B Chaires
Jul 5, 2008·Nature Protocols·Brandon T RuotoloCarol V Robinson
May 12, 2009·Drug Discovery Today·Gianni Chessari, Andrew J Woodhead
Mar 2, 2010·Pharmacological Research : the Official Journal of the Italian Pharmacological Society·Hongmei CaoRichard B van Breemen
Aug 20, 2010·Journal of Medicinal Chemistry·Christopher W MurrayAlison J-A Woolford
Oct 5, 2010·Current Opinion in Structural Biology·Francisco E TorresGlyn Williams
Jan 5, 2011·Analytical Chemistry·Yongsoo ChoiRichard R van Breemen
Mar 1, 2011·Journal of Pharmaceutical and Biomedical Analysis·Ranjith MuniguntiAngela I Calderón
Jul 14, 2011·Analytical and Bioanalytical Chemistry·Sayaka ShibataErkang Fan
Jun 2, 2012·Combinatorial Chemistry & High Throughput Screening·Charles E WhitehurstElliott Nickbarg
Sep 18, 2012·Journal of Biomolecular Screening·Minh-Dao Duong-ThiSten Ohlson
Jul 23, 2013·Proceedings of the National Academy of Sciences of the United States of America·Hernani Leonardo SilvestreAlessio Ciulli

❮ Previous
Next ❯

Citations

Sep 27, 2016·Journal of Pharmaceutical and Biomedical Analysis·Han WeiGuanghua Wu
Oct 8, 2017·Essays in Biochemistry·Daniel Shiu-Hin ChanChris Abell
Mar 27, 2018·Biophysical Reports·Yao PengZhi-Jie Liu
Nov 10, 2017·Essays in Biochemistry·Rob L M van Montfort, Paul Workman
Jul 25, 2018·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Yingjie HeHongqi Xie
Jan 9, 2021·Signal Transduction and Targeted Therapy·Dehua YangMing-Wei Wang
Aug 28, 2021·International Journal of Molecular Sciences·Hun Hwan KimGon Sup Kim

❮ Previous
Next ❯

Related Concepts

Related Feeds

Antivirals (ASM)

Antivirals are medications that are used specifically for treating viral infections. Discover the latest research on antivirals here.

Antivirals

Antivirals are medications that are used specifically for treating viral infections. Discover the latest research on antivirals here.