High-Resolution and High-Throughput Protocols for Measuring Drug/Human Serum Albumin Interactions Using BIACORE

Rich, Rebecca L.; Day, Yasmina S.N.; Morton, Thomas A.; Myszka, David G.

doi:10.1006/abio.2001.5314

Cited by 243 publications

(200 citation statements)

References 53 publications

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“…advanced to the point where it is possible to measure directly small molecules interacting with immobilized macromolecular targets (1,2). This development suggests that biosensor analysis will become an important secondary screening tool in drug discovery, confirming hits from primary screens and providing detailed kinetics for lead optimization (3,4).…”

Section: S Urface Plasmon Resonance (Spr) Biosensor Technology Hasmentioning

confidence: 99%

Kinetic analysis of estrogen receptor/ligand interactions

Rich

Hoth²,

Geoghegan³

et al. 2002

Proc. Natl. Acad. Sci. U.S.A.

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209

126

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Surface plasmon resonance biosensor technology was used to directly measure the binding interactions of small molecules to the ligand-binding domain of human estrogen receptor. In a screening mode, specific ligands of the receptor were easily discerned from nonligands. In a high-resolution mode, the association and dissociation phase binding responses were shown to be reproducible and could be fit globally to a simple interaction model to extract reaction rate constants. On average, antagonist ligands (such as tamoxifen and nafoxidine) were observed to bind to the receptor with association rates that were 500-fold slower than agonists (such as estriol and ␤-estradiol). This finding is consistent with these antagonists binding to an altered conformation of the receptor. The biosensor assay also could identify subtle differences in how the same ligand interacted with two different isoforms of the receptor (␣ and ␤). The biosensor's ability to determine kinetic rate constants for small molecule͞protein interactions provides unique opportunities to understand the mechanisms associated with complex formation as well as new information to drive the optimization of drug candidates.S urface plasmon resonance (SPR) biosensor technology has advanced to the point where it is possible to measure directly small molecules interacting with immobilized macromolecular targets (1, 2). This development suggests that biosensor analysis will become an important secondary screening tool in drug discovery, confirming hits from primary screens and providing detailed kinetics for lead optimization (3, 4). To illustrate the utility of current SPR technology, the binding properties of small compounds (200-500 Da) interacting with human estrogen receptor (ER) were analyzed.Ligand binding to ER is responsible for controlling the basic biology of estrogen-sensitive tissues. Using selective agonists or antagonists to modulate this biology is the focus of significant activity in the pharmaceutical industry (5-9). To date, a ligand's binding properties for ER have mainly been determined by equilibrium binding assays that often employ radiolabeled compounds and require overnight incubations. Here, we demonstrate how optical biosensors may be used to determine both kinetic and equilibrium binding constants for compounds interacting with ER in real time without labeling either binding partner.SPR biosensor experiments require immobilizing one reactant on a surface and monitoring its binding to a second reactant in solution. An antibody-capturing method was used to study the dynamics of ER͞ligand interactions. This assay format created a chemically homogenous receptor surface and allowed us to determine rapidly the binding properties of a variety of compounds. We examined the binding of 12 compounds (shown in Fig. 1) having differing receptor activities: both estrogen and non-estrogen agonists, SERMs (selective ER modulators, which for this discussion are referred to as antagonists), and nonbinding control compounds that possess core structures sim...

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Section: S Urface Plasmon Resonance (Spr) Biosensor Technology Hasmentioning

confidence: 99%

Kinetic analysis of estrogen receptor/ligand interactions

Rich

Hoth²,

Geoghegan³

et al. 2002

Proc. Natl. Acad. Sci. U.S.A.

Self Cite

209

126

View full text Add to dashboard Cite

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“…In the mass spectrum of the other cluster of ion peaks, four charge states ranging from 15ϩ to 19ϩ were observed, corresponding to HSA in its native state [44,45]. At this state, the HSA can bind effectively with ligands to form noncovalent complexes, and such complex formation between WAR and HSA with different protein-ligand ratios has been reported [46,47].…”

Section: Noncovalent Interaction Between Hsa and Warmentioning

confidence: 96%

Effect of tanshinone IIA on the noncovalent interaction between warfarin and human serum albumin studied by electrospray ionization mass spectrometry

Liu

Wang

Cai

et al. 2008

J. Am. Soc. Mass Spectrom.

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Enhanced anticoagulation and/or even bleeding are often observed when patients on long-term warfarin (WAR) therapy consumed Danshen, a well-known medicinal herb in traditional Chinese medicine (TCM). This study demonstrates that altered WAR metabolism, arising from its interaction with the active components in Danshen, played a significant role in this curative effect. Mass spectrometric techniques including ESI-ITMS (electrospray ionization ion-trap mass spectrometry) and ESI-TOF (time-of-flight)-MS have been developed for the study of such drug-herb interactions. The experimental approach involved a detailed analysis and comparison of WAR metabolites in vivo from blood or urine of rats that had been orally administrated with WAR, either singly or together with the representative bioactive component of Danshen-lipid soluble TIIA (Tanshinon IIA), and a study of the interaction of human serum albumin (HSA), WAR, and water-soluble sodium tanshinone IIA sulfonate (STS) in vitro. Results demonstrate that TIIA accelerates the metabolic rate of WAR, whereas STS displaces WAR from the WAR-HSA complex, resulting in an increase of free WAR concentration in blood. It is suggested that the elevated level and enhanced metabolism of WAR is responsible for the over-anticoagulation effect observed. (J Am Soc Mass Spectrom 2008, 19, 1568 -1575

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“…This methodology results particularly promising for their potential application in drug protein binding screening and in kinetics investigations. [22][23][24][25][26] The present review essentially focuses on the application of the use of both immobilized albumin (biochromatography) and CD on the same protein-drug system in solution, largely on the basis of our previous publications. These methodologies have been shown to be efficient in characterizing the binding sites on the protein and to give evidence of the changes in the binding properties of the protein arising by interaction between different ligands.…”

Section: Current Approaches For the Study Of The Binding Properties Omentioning

confidence: 99%

“…This methodology is well suited for screening assay, because of the miniaturization of the system and the short time for the analysis that facilitates its automatization. [22][23][24][25][26] Moreover, with optical biosensors, neither of the interacting molecules needs to be labeled, and the response obtained is directly proportional to the mass of the analyte that binds to the surface. Therefore, the ligand binding interaction occurs in a condition as close as possible to its native state.…”

Section: Biorecognition On Surface Immobilized Hsamentioning

confidence: 99%

Drug binding to human serum albumin: Abridged review of results obtained with high‐performance liquid chromatography and circular dichroism

2006

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The drug binding to plasma and tissue proteins are fundamental factors in determining the overall pharmacological activity of a drug. Human serum albumin (HSA), together with a 1 -acid glycoprotein (AGP), are the most important plasma proteins, which act as drug carriers, with drug pharmacokinetic implications, resulting in important clinical impacts for drugs that have a relatively narrow therapeutic index. This review focuses on the combination of biochromatography and circular dichroism as an effective approach for the characterization of albumin binding sites and their enantioselectivity. Furthermore, their applications to the study of changes in the binding properties of the protein arising by the reversible or covalent binding of drugs are discussed, and examples of physiological relevance reported. Perspectives of these studies reside in supporting the development of new drugs, which require miniaturization to facilitate the screening of classes of compounds for their binding to the target protein, and a deeper characterization of the mechanisms involved in the molecular recognition processes.

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High-Resolution and High-Throughput Protocols for Measuring Drug/Human Serum Albumin Interactions Using BIACORE

Cited by 243 publications

References 53 publications

Kinetic analysis of estrogen receptor/ligand interactions

Kinetic analysis of estrogen receptor/ligand interactions

Effect of tanshinone IIA on the noncovalent interaction between warfarin and human serum albumin studied by electrospray ionization mass spectrometry

Drug binding to human serum albumin: Abridged review of results obtained with high‐performance liquid chromatography and circular dichroism

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