Sensing Discrete Streptavidin-Biotin Interactions with Atomic Force Microscopy

G, Lee; Kidwell, David A.; Colton, Richard J.

doi:10.1021/la00014a003

Cited by 728 publications

(578 citation statements)

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“…In these experiments the cantilever deflection signal, measured in nAmp, was converted to a deflection distance (nm) using the gradient of the retract trace in the contact region of the force curve (see Fig. 1) [8]. This distance, d, was converted to the force acting on the AFM probe (nN) using the cantilever spring constant (k), and Hooke's law (F=-kd).…”

Section: Spm Analysismentioning

confidence: 99%

“…By attaching complementary bionolecules to the AFM probe and the opposing surface this lbility has been exploited to measure the forces required to ~eparate specific biomolecular interactions [8][9][10][11][12][13][14][15]. The strepavidin-biotin complex as a model receptor-ligand interaction 1as generated much interest by virtue of its high specificity md affinity (K~ = 10 -15 mol 1-1) and general applicability as an immobilisation method [8,13,16,17].…”

Section: Introductionmentioning

confidence: 99%

“…The long and short range forces controlling this specific interaction have been well characterised [16,17] and this combined with the availability of structural [21] and thermodynamic [22,23] data has made it an ideal system to study receptor-ligand interactions. Lee et al [8] used biotin fuctionalised glass beads attached to AFM cantilevers and streptavidin coated mica surfaces to estimate the strength of a single streptavidin-biotin bond. A similar approach was adopted by Gaub et al [9,10,12,13], who measured quantised forces of interaction between a biotinylated agarose bead and an AFM probe functionalised with avidin, a 67 kDa protein closely related to streptavidin.…”

Section: Introductionmentioning

confidence: 99%

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In situ observation of streptavidin‐biotin binding on an immunoassay well surface using an atomic force microscope

et al. 1996

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Polystyrene microtitre wells are commonly used as supports for the enzyme-linked immunosorbent assay (ELISA) method of biomolecular detection, which is employed in the routine diagnosis of a variety of medical conditions. We have used an atomic force microscope (AFM) to directly monitor specific molecular interactions between individual streptavidin and biotin molecules on such wells. This was achieved by functionalising an AFM probe with biotin and monitoring the adhesive forces between the probe and a streptavidin coated immunoassay well. The results demonstrate that the AFM may be employed as an analytical tool to study the interactions between biomolecules involved in immunoassay systems.

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Section: Spm Analysismentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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In situ observation of streptavidin‐biotin binding on an immunoassay well surface using an atomic force microscope

et al. 1996

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“…Dès 1994, deux équipes de chercheurs [19,20] observaient la rupture de liaisons moléculaires individuelles au moyen d'une méthode entièrement différente: la microscopie de force atomique. Cette approche puissante repose sur un concept extrêmement simple ( Figure 2C, D): une surface couverte de ligands est approchée d'une pointe extrêmement fine (un rayon de courbure de quelques dizaines de nanomètres) recouverte de récepteurs.…”

Section: Les Sélectinesunclassified

“…Le modèle de l'interaction avidine/biotine avait été choisi parce qu'il s'agit de la plus forte interaction non covalente caractérisée en biologie. La force de rupture mesurée était comprise entre 160 pN [19] et 300-400 pN [20]. Ces variations pouvaient-elles s'expliquer par les incertitudes expérimentales ?…”

Section: Les Sélectinesunclassified

Liaisons « accrocheuses » et mécanique moléculaire

2003

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Microscopy for recognition of individual biomolecules

Schmidt

Hinterdorfer

Schindler

1999

Microsc. Res. Tech.

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One frontier challenge in microscopy and analytical chemistry is the analysis of soft matter at the single molecule level with biological systems as most complex examples. Towards this goal we have developed two novel microscopy methods. Both employ highly specific molecular recognition schemes used by nature-the recognition of specific protein sites by antibodies and ligands. One method uses fluorescence labeled ligands for detecting single molecules in fluid systems like membranes (Fig. 1B). Unitary signals are reliably resolved even for millisecond illumination periods. The knowledge of the unitary signal from single molecules permits the determination of stoichiometries of component association (Fig. 3). Direct imaging of the diffusional path of single molecules became possible for the first time (Fig. 4). Using linear polarized excitation, the angular orientation of single molecules can be analyzed (single molecule linear dichroism, (Fig. 5), which opens a new perspective for detecting conformational changes of single biomolecules. In the other method, an antibody is flexibly linked to the tip of an atomic-force microscope. This permits the identification of receptors in multi-component systems. Molecular mapping of biosurfaces and the study of molecular dynamics in the ms to s range become possible with atomic force microscopy.

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Sensing Discrete Streptavidin-Biotin Interactions with Atomic Force Microscopy

Cited by 728 publications

References 1 publication

In situ observation of streptavidin‐biotin binding on an immunoassay well surface using an atomic force microscope

In situ observation of streptavidin‐biotin binding on an immunoassay well surface using an atomic force microscope

Liaisons « accrocheuses » et mécanique moléculaire

Microscopy for recognition of individual biomolecules

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