SAW Sensor System for Marker‐Free Molecular Interaction Analysis

Perpeet, M.; Glass, Samuel V.; Gronewold, Thomas M. A.; Kiwitz, A.; Malavé, A.; Stoyanov, Ivan; Tewes, M.; Quandt, Eckhard

doi:10.1080/000327106006714063

Cited by 44 publications

(45 citation statements)

References 11 publications

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“…The increase in mass on the biosensor surface upon drug binding induces a phase shift of the acoustic wave, whereas changes in the viscoelasticity on the surface -e.g. by a conformational change of the immobilized protein -mainly affect its amplitude [9,10]. While kinetic-binding analysis of small molecules by SPR is nowadays performed in a routine fashion for soluble protein targets like proteases or kinases, it is still challenging for integral membrane proteins like GPCRs.…”

Section: Contents Lists Available At Sciencedirectmentioning

confidence: 99%

Label-free detection of small-molecule binding to a GPCR in the membrane environment

Heym

Hornberger

Lakics

et al. 2015

Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics

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Section: Contents Lists Available At Sciencedirectmentioning

confidence: 99%

Label-free detection of small-molecule binding to a GPCR in the membrane environment

Heym

Hornberger

Lakics

et al. 2015

Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics

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“…A recently-explored alternative to SPR is the surface acoustic wave (SAW) technology in which the piezoelectric effect of mass differences is employed for bioaffinity detection [6 -9]. SAW is now becoming increasingly important for the study of biomacromolecular interactions due to its high detection sensitivity in dilute solutions.Advantages of SAW in comparison to classical immuno-analytical techniques are the direct and rapid determination of association/dissociation constants with small sample amounts, and without labeling approaches or recalibration for buffer changes being required [6]. While providing sensitive and accurate determinations of binding/dissociation constants (K i or K D ), a major limitation of all bioaffinity methods is the lack of direct identification of the affinity-bound ligands.…”

mentioning

confidence: 99%

“…Advantages of SAW in comparison to classical immuno-analytical techniques are the direct and rapid determination of association/dissociation constants with small sample amounts, and without labeling approaches or recalibration for buffer changes being required [6]. While providing sensitive and accurate determinations of binding/dissociation constants (K i or K D ), a major limitation of all bioaffinity methods is the lack of direct identification of the affinity-bound ligands.…”

mentioning

confidence: 99%

On-line bioaffinity-electrospray mass spectrometry for simultaneous detection, identification, and quantification of protein-ligand interactions

Drăguşanu

Petre

Slamnoiu

et al. 2010

J. Am. Soc. Mass Spectrom.

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We describe here an on-line combination of a surface acoustic wave (SAW) biosensor with electrospray ionization mass spectrometry (SAW-ESI-MS) that enables the direct detection, identification, and quantification of affinity-bound ligands from a protein-ligand complex on a biosensor chip. A trapping column was used between the SAW-biosensor and the electrospray mass spectrometer equipped with a micro-guard column, which provides simultaneous sample concentration and desalting for the mass spectrometric analysis of the dissociated ligand. First applications of the on-line SAW-ESI-MS combination include (1), differentiation of ␤-amyloid (A␤) epitope peptides bound to anti-A␤ antibodies; (2), the identification of immobilized Substance P peptide-calmodulin complex; (3), identification and quantification of the interaction of 3-nitrotyrosine-modified peptides with nitrotyrosine-specific antibodies; and (4), identification of immobilized anti-␣-synuclein-human ␣-synuclein complex. Quantitative determinations of protein-ligand complexes by SAW yielded dissociation constants (K D ) from micro-to low nanomolar sample concentrations. The on-line bioaffinity-ESI-MS combination presented here is expected to enable broad bioanalytical application to the simultaneous, label-free determination and quantification of biopolymer-ligand interactions, as diverse as antigen-antibody and lectin-carbohydrate complexes. (J Am Soc Mass Spectrom 2010, 21, 1643-1648 [3,4]. In particular, SPR has developed into an efficient tool for analysis of biomolecular recognition processes at a biosensor surface, and has been applied to the quantification of a variety of biopolymer interactions [4,5]. A recently-explored alternative to SPR is the surface acoustic wave (SAW) technology in which the piezoelectric effect of mass differences is employed for bioaffinity detection [6 -9]. SAW is now becoming increasingly important for the study of biomacromolecular interactions due to its high detection sensitivity in dilute solutions.Advantages of SAW in comparison to classical immuno-analytical techniques are the direct and rapid determination of association/dissociation constants with small sample amounts, and without labeling approaches or recalibration for buffer changes being required [6]. While providing sensitive and accurate determinations of binding/dissociation constants (K i or K D ), a major limitation of all bioaffinity methods is the lack of direct identification of the affinity-bound ligands. In contrast, the combination of biosensor detection and mass spectrometry enables both identification and quantification of bioaffinity interactions of biopolymers. Here we describe an on-line combination of an SAW biosensor with electrospray ionization mass spectrometry, SAW-ESI-MS. The on-line coupling between the SAW-sensor chip and the ESI-MS source was achieved by a incorporating a standard trapping column setup, using a six-port valve, a guard column, and a micropump system. The six-port valve interface provides both ligand concentration and an...

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“…For preparation of the sensor, an AT cut quartz crystal was processed by photolithography as described previously (Perpeet et al, 2006). The obtained sensor chips of Love-wave sensor type with five sensor elements ( Fig.…”

Section: Love-wave Sensor Set-up and Immobilization Of Aptamersmentioning

confidence: 99%

“…antibodies or aptamers. Previously, a S-sens ® K5 biosensor with a detection limit of <1 pg/mm 2 (Perpeet et al, 2006;Schlensog et al, 2004) was used to characterize human thrombin, a serine protease acting as a component of the blood-coagulation cascade and by use of a sensor-immobilized anti-thrombin RNA-aptamer (White et al, 2001). The main role of thrombin as the last protease in the clotting cascade (Holland et al, 2000) is to process numerous enzymatic actions on other blood proteins and on receptors (Coughlin, 2005).…”

Section: Introductionmentioning

confidence: 99%