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Electrospray ionization mass spectrometry as a tool for determination of drug binding sites to human serum albumin by noncovalent interaction
KTH, School of Chemical Science and Engineering (CHE), Chemistry, Analytical Chemistry.
Biovitrum AB, Dept Analyt Sci.
KTH, School of Chemical Science and Engineering (CHE), Chemistry, Analytical Chemistry.
2005 (English)In: Rapid Communications in Mass Spectrometry, ISSN 0951-4198, E-ISSN 1097-0231, Vol. 19, no 12, 1637-1643 p.Article in journal (Refereed) Published
Abstract [en]

Most proteins in blood plasma bind ligands. Human serum albumin (HSA) is the main transport protein with a very high capacity for binding of endogenous and exogenous compounds in plasma. Many pharmacokinetic properties of a drug depend on the level of binding to plasma proteins. This work reports studies of noncovalent interactions by means of nanoelectrospray ionization mass spectrometry (nanoESI-MS) for determination of the specific binding of selected drug candidates to HSA. Warfarin, iopanoic acid and digitoxin were chosen as site-specific probes that bind to the main sites of HSA. Two drug candidates and two known binders to HSA were analyzed using a competitive approach. The drugs were incubated with the target protein followed by addition of site-specific probes, one at a time. The drug candidates showed predominant affinity to site I (warfarin site). Naproxen and glyburide showed affinity to both sites I and II. The advantages of nanoE-SI-MS for these studies are the sensitivity, the absence of labeled molecules and the short method development time.

Place, publisher, year, edition, pages
2005. Vol. 19, no 12, 1637-1643 p.
Keyword [en]
PROTEIN-LIGAND COMPLEXES; ULTRAFILTRATION; COMPETITION; RESOLUTION; AFFINITY; ACID
National Category
Analytical Chemistry
Identifiers
URN: urn:nbn:se:kth:diva-8360DOI: 10.1002/rcm.1967ISI: 000229846800008Scopus ID: 2-s2.0-20644463474OAI: oai:DiVA.org:kth-8360DiVA: diva2:13661
Note
QC 20100705Available from: 2008-05-07 Created: 2008-05-07 Last updated: 2017-12-14Bibliographically approved
In thesis
1. Electrospray Ionization Mass Spectrometry for Determination of Noncovalent Interactions in Drug Discovery
Open this publication in new window or tab >>Electrospray Ionization Mass Spectrometry for Determination of Noncovalent Interactions in Drug Discovery
2008 (English)Doctoral thesis, comprehensive summary (Other scientific)
Abstract [en]

Noncovalent interactions are involved in many biological processes in which biomolecules bind specifically and reversibly to a partner. Often, proteins do not have a biological activity without the presence of a partner, a ligand. Biological signals are produced when proteins interact with other proteins, peptides, oligonucleotides, nucleic acids, lipids, metal ions, polysaccharides or small organic molecules. Some key steps in the drug discovery process are based on noncovalent interactions. We have focused our research on the steps involving ligand screening, competitive binding and ‘off-target’ binding. The first paper in this thesis investigated the complicated electrospray ionization process with regards to noncovalent complexes. We have proposed a model that may explain how the equilibrium between a protein and ligand changes during the droplet evaporation/ionization process.

The second paper describes an evaluation of an automated chip-based nano-ESI platform for ligand screening. The technique was compared with a previously reported method based on nuclear magnetic resonance (NMR), and excellent correlation was obtained between the results obtained with the two methods. As a general conclusion we believe that the automated nano-ESI/MS should have a great potential to serve as a complementary screening method to conventional HTS. Alternatively, it could be used as a first screening method in an early phase of drug development programs when only small amounts of purified targets are available.

In the third article, the advantage of using on-line microdialysis as a tool for enhanced resolution and sensitivity during detection of noncovalent interactions and competitive binding studies by ESI-MS was demonstrated. The microdialysis device was improved and a new approach for competitive binding studies was developed.

The last article in the thesis reports studies of noncovalent interactions by means of nanoelectrospray ionization mass spectrometry (nanoESI-MS) for determination of the specific binding of selected drug candidates to HSA. Two drug candidates and two known binders to HSA were analyzed using a competitive approach. The drugs were incubated with the target protein followed by addition of site-specific probes, one at a time. The drug candidates showed predominant affinity to site I (warfarin site). Naproxen and glyburide showed affinity to both sites I and II.

Place, publisher, year, edition, pages
Stockholm: KTH, 2008. viii, 60 p.
Series
Trita-CHE-Report, ISSN 1654-1081 ; 2008:33
Keyword
Mass spectrometry, electrospray ionization, drug discovery, noncovalent interaction, complexes, human serum albumin (HSA), fatty acid binding protein (FABP), ribonuclease, ligand screening
National Category
Analytical Chemistry
Identifiers
urn:nbn:se:kth:diva-4730 (URN)978-91-7178-947-1 (ISBN)
Public defence
2008-05-23, E2, KTH, Lindstedtsvägen 3, Stockholm, 00:00
Opponent
Supervisors
Note
QC 20100705Available from: 2008-05-07 Created: 2008-05-07 Last updated: 2010-07-05Bibliographically approved

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