Methods of enumerating receptor molecules for specific binding partners on formed bodies and in solution

摘要

This invention relates generally to using fluorescent markers (labelled antibodies) and flow cytometry to enumerate the average number of receptors (antigen) on formed bodies (cells) in whole blood, and to evaluate the specific binding constant of the marker for the particular receptors. Mean channel fluorescence intensities of equilibrated marker- cell suspension mixtures, total concentrations of marker, and targeted cell counts obtained by established procedures are used to complete the analyses. The invention further describes a competitive binding assay between marker and unlabelled antibody for receptors shed from the surface of formed bodies into solution and receptors on the formed bodies to analyze for receptors in solution. Moreover, competitive binding assays between marker and unlabelled antibody for receptors on formed bodies in whole blood are described for determination of relative, specific, and average binding constants of unlabelled/labelled antibody for targeted receptors.