Detection of Antigen−Antibody Binding Events with the Atomic Force Microscope

Allen, Stephanie; Chen, Xinyong; Davies, John A.; Davies, Martyn C.; Dawkes, Adrian; Edwards, John C.; Roberts, Clive J.; Sefton, J.; Tendler, Saul J. B.; Williams, Philip M.

doi:10.1021/bi962531z

Cited by 331 publications

(266 citation statements)

References 26 publications

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“…4. After the postulation by Allen et al (16), these forces could represent the binding of multiple WNV E DIII protein and ␣V␤3 integrin pairs. The forces measured in Fig.…”

Section: Measurement Of Wnv E DIII Protein-␣v␤3 Integrin Binding Force-mentioning

confidence: 99%

“…This is in addition to providing many successful surface topographical information in biological studies (9 -15). It can also be used to study interactions between two molecules, such as antigens and their corresponding antibodies (16).…”

mentioning

confidence: 99%

“…On the retract phase of the force measurement, the maximum cantilever deflection is "related directly to the magnitude of the force required" to break the bonds holding the two interacting partners (16). The quantitative amount of force required to separate individual receptor-ligand interactions may be calculated from the generated force curve.…”

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confidence: 99%

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Quantifying the Specific Binding between West Nile Virus Envelope Domain III Protein and the Cellular Receptor αVβ3 Integrin

Lee¹,

Chu²,

2006

Journal of Biological Chemistry

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A previous study has illustrated that the ␣V␤3 integrin served as the functional receptor for West Nile virus (WNV) entry into cells. Domain III (DIII) of WNV envelope protein (E) was postulated to mediate virus binding to the cellular receptor. In this study, the specificity and affinity binding of WNV E DIII protein to ␣V␤3 integrin was confirmed with co-immunoprecipitation and receptor competition assay. Binding of WNV E DIII protein to ␣V␤3 integrin induced the phosphorylation of focal adhesion kinase that is required to mediate ligand-receptor internalization into cells. A novel platform was then developed using the atomic force microscopy to measure this specific binding force between WNV E DIII protein and the cellular receptor, ␣V␤3 integrin. The single protein pair-interacting force measured was in the range of 45 ؎ 5 piconewtons. This interacting force was highly specific as minimal force was measured in the WNV E DIII protein interaction with ␣V␤5 integrin molecules and heparan sulfate. These experiments provided an insight to quantitate virus-receptor interaction. Force measurement using atomic force microscopy can serve to quantitatively analyze the effect of candidate drugs that modulate virus-host receptor affinity.The family Flaviviridae is positive-sense, single-stranded RNA viruses that replicate in the cytoplasm of infected cells. Many members of the three genera (Flavivirus, Pestivirus, and Hepacivirus) belonging to this family are medically important human pathogens. West Nile virus (WNV), 4 an arthropod-transmitted Flavivirus, is the causative agent of the disease syndrome named West Nile fever including a spectrum of associated complication (meningoencephalitis) (1). This is a re-emerging arthropod-borne disease that is responsible for recent large outbreaks in the Western hemisphere. In 2004 there were 2313 human infections and 79 deaths reported in the United States (2). Currently, there is no vaccine or antiviral agent against this pathogenic virus.Crystallography data on the ectodomain of the Flavivirus E protein reveals three distinct domains: a central domain designated as domain I (DI), an elongated dimerization region designated as domain II (DII), and domain III (DIII), having an immunoglobulin-like constant domain (3). Both DII and DIII of the E protein have been suggested to be important for binding to the cellular receptor (4 -7). Chu and Ng (8) have recently documented the involvement of ␣V␤3 integrin in mediating the infectious entry of WNV into host cells. Specific binding between WNV and ␣V␤3 integrin can be illustrated by functional blocking antibodies against ␣V␤3 integrin and its subunits. The high antagonistic effect of recombinant WNV DIII protein on WNV infection in both mammalian and mosquito cells has also strongly suggested that DIII of WNV E protein functions as the receptor binding domain and is responsible for the recognition and attachment to the cellular receptor (7).In this study the specific interaction between the DIII of WNV E protein and its cellular rece...

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“…4. After the postulation by Allen et al (16), these forces could represent the binding of multiple WNV E DIII protein and ␣V␤3 integrin pairs. The forces measured in Fig.…”

Section: Measurement Of Wnv E DIII Protein-␣v␤3 Integrin Binding Force-mentioning

confidence: 99%

mentioning

confidence: 99%

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Quantifying the Specific Binding between West Nile Virus Envelope Domain III Protein and the Cellular Receptor αVβ3 Integrin

Lee¹,

Chu²,

2006

Journal of Biological Chemistry

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“…In this context, an interesting approach is to mix long chain alkanethiols with COOH terminal functions in a matrix of shorter OH-terminated alkanethiols, which ensures a certain mobility of the attached biomolecules and minimizes non-specific adsorption [16]. A third approach uses amine-functionalization procedures to covalently anchor biomolecules on silicon tips [17,18]. The amino-terminated surfaces are reacted with a cross-linker which provides the ligands with motional freedom and prevents their denaturation.…”

Section: Functionalized Tipsmentioning

confidence: 99%

Recent progress in AFM molecular recognition studies

Dufrêne

Hinterdorfer

2007

Pflugers Arch - Eur J Physiol

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During the past decade, remarkable advances have been made in using atomic force microscopy (AFM) for measuring the forces and the dynamics of the interaction between individual ligands and receptors, providing fundamental insights into molecular recognition processes. In addition, affinity imaging using either adhesion force mapping or dynamic recognition force mapping has offered a means to localize specific binding sites on model and cellular surfaces. These single-molecule analyses provide novel insight into the structure-function relationships of molecular recognition systems. In this review, we describe the principles of molecular recognition studies using AFM and provide a flavor of recent progress made in the field.

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“…9 Among those immunosensors, the capacitive immunosensor has attracted more interest with high sensitivity, rapid test time and common instrumentations [10][11][12][13][14][15] compared with other types of direct biosensors, such as surface plasmon resonance, 16,17 quartz crystal microbalance (QCM), 18 surface acoustic wave, 19 and surface scanning detection. 20 The fabrication of a biocompatible layer is more pivotal to a capacitive immunosensor. It must firstly be sufficiently insulated and hole-free, or else electro-activated ions can move through the layer, causing interference of the faradaic current, leading to a decrease or disappearance of the signal.…”

Section: Introductionmentioning

confidence: 99%