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

Research output: Contribution to journal › Article › peer-review

14 Citations (Scopus)

Abstract

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.

Original language	English
Pages (from-to)	794-796
Number of pages	3
Journal	Analytical Biochemistry
Volume	421
Issue number	2
DOIs	https://doi.org/10.1016/j.ab.2011.12.011
Publication status	Published - 15 Feb 2012
Externally published	Yes

Keywords

Kinetics
Label free
Protein interaction analysis
Rate and dissociation equilibrium constants
SPR imaging

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10.1016/j.ab.2011.12.011

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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.\}",

year = "2012",

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doi = "10.1016/j.ab.2011.12.011",

language = "English",

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journal = "Analytical Biochemistry",

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T1 - Method for estimating the single molecular affinity

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

Y1 - 2012/2/15

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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DO - 10.1016/j.ab.2011.12.011

M3 - Article

C2 - 22209736

AN - SCOPUS:84859714260

SN - 0003-2697

VL - 421

SP - 794

EP - 796

JO - Analytical Biochemistry

JF - Analytical Biochemistry

IS - 2

ER -

Method for estimating the single molecular affinity

Abstract

Keywords

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