These tools will no longer be maintained as of December 31, 2024. Archived website can be found here. PubMed4Hh GitHub repository can be found here. Contact NLM Customer Service if you have questions.


Pubmed for Handhelds

PUBMED FOR HANDHELDS

Search MEDLINE/PubMed

  • Title: Direct measurement of antibody affinity distribution by hapten-inhibition enzyme immunoassay.
    Author: Nieto A, Gaya A, Jansa M, Moreno C, Vives J.
    Journal: Mol Immunol; 1984 Jun; 21(6):537-43. PubMed ID: 6379416.
    Abstract:
    A rapid, simple and reliable technique for determining the affinities of antibody subpopulations in a complex mixture is described. The principle of this method is that antigen conc can be represented as the amount of antigen immobilized on the polystyrene surface of a microwell containing a fixed vol of diluted antibody. Thus, by measuring the proportion of antibody bound to different wells coated with varying amounts of antigen, it is a straightforward matter to calculate an affinity distribution. We have verified that: (1) the amount of antigen bound to a polystyrene plate is proportional to the concn of antigen used for sensitization and follows a typical saturation curve; (2) the antibodies bound to plates sensitized with low concns of antigen are of higher affinity than those bound to plates sensitized with high concns of antigen; (3) an apparent affinity constant (aK) is defined as the reciprocal concn of free hapten required for 50% inhibition of antibody binding to immobilized antigen; (4) the aK determined by this method is in close agreement with the intrinsic affinity constant (K) measured by fluorescence quenching or the Farr assay; and (5) that during the course of immunization in vivo there is a clear shift to higher-affinity antibody subpopulations.
    [Abstract] [Full Text] [Related] [New Search]