Details

Autor(en) / Beteiligte

• Rabbany, Sina Y.
• Kusterbeck, Anne W.
• Bredehorst, Reinhard
• Ligler, Frances S.

Titel

Effect of antibody density on the displacement kinetics of a flow immunoassay

Ist Teil von

• Journal of immunological methods, 1994-02, Vol.168 (2), p.227-234

Ort / Verlag

Amsterdam: Elsevier B.V

Erscheinungsjahr

1994

Quelle

Access via ScienceDirect (Elsevier)

Beschreibungen/Notizen

• This study investigates the effect of antibody density on the kinetics of a solid-phase displacement immunoassay. Conducted in flow under nonequilibrium conditions, the assay utilizes a monoclonal antibody to the cocaine metabolite benzoylecgonine, which has been immobilized onto Sepharose beads and saturated with fluorophorelabeled antigen. Displacement of antibody-bound labeled antigen by non-labeled antigen occurs when sample is introduced in the buffer flow. Comparison of matrices coated with two different antibody densities revealed that the displacement efficiency is a function of the density of antibody-bound labeled antigen. A higher density of antibody provides a higher amount of displaced labeled antigen, but the displacement efficiency of the assay is decreased. The effect of antibody density on the immunoassay kinetics was analyzed using a mathematical formulation developed to characterize antibody-antigen interactions at solid-liquid interfaces. Higher antibody density proved to be associated with a lower apparent dissociation rate constant. The implications of these results on the design of immunassays in flow are discussed.