Molina-Bolívar J A, Galisteo-González F, Hidalgo-Alvarez R
Department of applied Physics, Faculty of Sciences, Campus Fuenteneuva, University of Granada, Spain.
J Immunol Methods. 1998 Feb 1;211(1-2):87-95. doi: 10.1016/s0022-1759(97)00189-0.
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.
