Affordable Access

Publisher Website

Measurement of antibody affinity for cell surface antigens using an enzyme-linked immunosorbent assay

Authors
Journal
Journal of Immunological Methods
0022-1759
Publisher
Elsevier
Publication Date
Volume
125
Identifiers
DOI: 10.1016/0022-1759(89)90090-2
Keywords
  • Affinity Constant
  • Monoclonal Antibody
  • Cell Surface Antigen
  • Elisa
  • Flow Cytometry
Disciplines
  • Biology

Abstract

Abstract We present a fast, simple, and accurate method to determine the affinity constants of antibodies that bind to cell surface antigens. This procedure utilizes intact cells and native, unmodified antibody in a conventional enzyme-linked immunosorbent assay. Target cells are incubated with serial dilutions of antibody and allowed to reach equilibrium. Cells are then pelleted by centrifugation, and aliquots of unbound antibody in the supernatant are added to a microtiter plate precoated with capture antibody and measured in a conventional enzyme-linked immunosorbent assay (ELISA). We measured the affinity constant of murine monoclonal antibody CLB-1H-gran2, which binds to K562 cells (a human erythroleukemia line), and compared the ELISA-based results to those obtained by flow cytometric determination of antibody affinity. The affinity constants obtained by the two methods are in good aggreement. The affinity constant is calculated utilizing only the concentrations of bound and free antibody, so that the actual antigen concentration (or number of antigenic sites per cell) need not be known. However, the number of antibody molecules bound per cell can be estimated from the results.

There are no comments yet on this publication. Be the first to share your thoughts.

Statistics

Seen <100 times
0 Comments

More articles like this

Measurement of antibody affinity for cell surface...

on Journal of Immunological Metho... Dec 20, 1989

Measurement of cell proliferation by enzyme-linked...

on Cell Proliferation September 1991

Direct measurement of antibody production in cell...

on Journal of Immunological Metho... Feb 11, 1985

Direct measurement of antibody production in cell...

on Journal of Immunological Metho... Jan 01, 1985
More articles like this..