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A rigorous multiple independent binding site model for determining cell-based equilibrium dissociation constants.
Drake, Andrew W; Klakamp, Scott L.
Afiliação
  • Drake AW; Global Dev. DMPK and Bioanalysis, Biotherapeutics San Francisco, AstraZeneca Pharmaceuticals LP, 24500 Clawiter Road, Hayward, CA 94545, USA.
J Immunol Methods ; 318(1-2): 147-52, 2007 Jan 10.
Article em En | MEDLINE | ID: mdl-17141800
A new 4-parameter nonlinear equation based on the standard multiple independent binding site model (MIBS) is presented for fitting cell-based ligand titration data in order to calculate the ligand/cell receptor equilibrium dissociation constant and the number of receptors/cell. The most commonly used linear (Scatchard Plot) or nonlinear 2-parameter model (a single binding site model found in commercial programs like Prism(R)) used for analysis of ligand/receptor binding data assumes only the K(D) influences the shape of the titration curve. We demonstrate using simulated data sets that, depending upon the cell surface receptor expression level, the number of cells titrated, and the magnitude of the K(D) being measured, this assumption of always being under K(D)-controlled conditions can be erroneous and can lead to unreliable estimates for the binding parameters. We also compare and contrast the fitting of simulated data sets to the commonly used cell-based binding equation versus our more rigorous 4-parameter nonlinear MIBS model. It is shown through these simulations that the new 4-parameter MIBS model, when used for cell-based titrations under optimal conditions, yields highly accurate estimates of all binding parameters and hence should be the preferred model to fit cell-based experimental nonlinear titration data.
Assuntos
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Base de dados: MEDLINE Assunto principal: Receptores de Superfície Celular / Ligantes / Modelos Químicos Idioma: En Ano de publicação: 2007 Tipo de documento: Article
Buscar no Google
Base de dados: MEDLINE Assunto principal: Receptores de Superfície Celular / Ligantes / Modelos Químicos Idioma: En Ano de publicação: 2007 Tipo de documento: Article