Method for estimating the single molecular affinity

Richard B.M. Schasfoort; Wim De Lau; Alex Van Der Kooi; Hans Clevers; Gerard H.M. Engbers

doi:10.1016/j.ab.2011.12.011

Method for estimating the single molecular affinity

Richard B.M. Schasfoort, Wim De Lau, Alex Van Der Kooi, Hans Clevers, Gerard H.M. Engbers

Onderzoeksoutput: Bijdrage aan tijdschrift › Artikel › peer review

13 Citaten (Scopus)

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Affinity constants (k_d, k_a, and K_D) can be determined by methods that apply immobilized ligands such as immunoassays and label-free biosensor technologies. This article outlines a new surface plasmon resonance (SPR) array imaging method that yields affinity constants that can be considered as the best estimate of the affinity constant for single biomolecular interactions. Calculated rate (k_d and k_a) and dissociation equilibrium (K_D) constants for various ligand densities and analyte concentrations are extrapolated to the K_D at the zero response level (K_D^R0). By applying this method to an LGR5-exo-Fc-RSPO1-FH interaction couple, the K_D^R0 was determined as 3.1 nM.

Originele taal-2	Engels
Pagina's (van-tot)	794-796
Aantal pagina's	3
Tijdschrift	Analytical Biochemistry
Volume	421
Nummer van het tijdschrift	2
DOI's	https://doi.org/10.1016/j.ab.2011.12.011
Status	Gepubliceerd - 15 feb. 2012
Extern gepubliceerd	Ja

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title = "Method for estimating the single molecular affinity",

abstract = "Affinity constants (kd, ka, and KD) can be determined by methods that apply immobilized ligands such as immunoassays and label-free biosensor technologies. This article outlines a new surface plasmon resonance (SPR) array imaging method that yields affinity constants that can be considered as the best estimate of the affinity constant for single biomolecular interactions. Calculated rate (kd and ka) and dissociation equilibrium (KD) constants for various ligand densities and analyte concentrations are extrapolated to the KD at the zero response level (KDR0). By applying this method to an LGR5-exo-Fc-RSPO1-FH interaction couple, the KDR0 was determined as 3.1 nM.",

keywords = "Kinetics, Label free, Protein interaction analysis, Rate and dissociation equilibrium constants, SPR imaging",

author = "Schasfoort, {Richard B.M.} and {De Lau}, Wim and {Van Der Kooi}, Alex and Hans Clevers and Engbers, {Gerard H.M.}",

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AU - Schasfoort, Richard B.M.

AU - De Lau, Wim

AU - Van Der Kooi, Alex

AU - Clevers, Hans

AU - Engbers, Gerard H.M.

PY - 2012/2/15

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N2 - Affinity constants (kd, ka, and KD) can be determined by methods that apply immobilized ligands such as immunoassays and label-free biosensor technologies. This article outlines a new surface plasmon resonance (SPR) array imaging method that yields affinity constants that can be considered as the best estimate of the affinity constant for single biomolecular interactions. Calculated rate (kd and ka) and dissociation equilibrium (KD) constants for various ligand densities and analyte concentrations are extrapolated to the KD at the zero response level (KDR0). By applying this method to an LGR5-exo-Fc-RSPO1-FH interaction couple, the KDR0 was determined as 3.1 nM.

AB - Affinity constants (kd, ka, and KD) can be determined by methods that apply immobilized ligands such as immunoassays and label-free biosensor technologies. This article outlines a new surface plasmon resonance (SPR) array imaging method that yields affinity constants that can be considered as the best estimate of the affinity constant for single biomolecular interactions. Calculated rate (kd and ka) and dissociation equilibrium (KD) constants for various ligand densities and analyte concentrations are extrapolated to the KD at the zero response level (KDR0). By applying this method to an LGR5-exo-Fc-RSPO1-FH interaction couple, the KDR0 was determined as 3.1 nM.

KW - Kinetics

KW - Label free

KW - Protein interaction analysis

KW - Rate and dissociation equilibrium constants

KW - SPR imaging

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JO - Analytical Biochemistry

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Method for estimating the single molecular affinity

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