Particle enhanced immunoassays stabilized by hydration forces: a comparative study between IgG and F(ab)2 immunoreactivity.
In previous publications we have discussed the stabilization mechanism of hydration forces as applied to the development of latex agglutination tests. We describe here how we have obtained stable and reactive IgG-latex conjugates in a high-ionic-strength reaction buffer. To this end we have made agglutination tests with polystyrene beads sensitized with IgG, measuring the immunoaggregation reaction with human C-reactive protein in a stopped-flow nephelometer. The results are compared to those obtained with a F(ab')2-latex conjugate with similar antibody molecule coverage. Adsorption isotherms of F(ab')2 and IgG on latex at pH 7.2 were obtained to study the affinity of these antibodies for the surface. The results of the electrokinetic characterization of the antibody-latex conjugates agree satisfactorily with those obtained from stability studies. This research throws light upon the use of hydration forces as a new approach to stabilizing immunoassay reagents that are colloidally unstable in physiological reaction buffers.
Molina-Bolívar
J A
JA
Department of applied Physics, Faculty of Sciences, Campus Fuenteneuva, University of Granada, Spain.
Galisteo-González
F
F
Hidalgo-Alvarez
R
R
D000327:Adsorption; D002097:C-Reactive Protein / Q000276:immunology; D003102:Colloids; D006801:Humans; D007140:Immunoglobulin Fab Fragments / Q000276:immunology*; D007074:Immunoglobulin G / Q000276:immunology*; D007841:Latex Fixation Tests; D011137:Polystyrenes
D003102:Colloids; D007140:Immunoglobulin Fab Fragments; D007074:Immunoglobulin G; D011137:Polystyrenes; C024075:styrofoam; D002097:C-Reactive Protein