Turbidimetric assay of IgG with use of single monoclonal antibodies.
We describe a turbidimetric assay for quantifying total immunoglobulin G (IgG) in serum with use of a single monoclonal antibody. The reaction, monitored by a centrifugal analyzer, is technically simple, rapid, and precise. Buffer of low ionic strength and polyethylene glycol are required for formation of detectable antibody-antigen complexes. We measured IgG concentrations in 49 polyclonal sera (Group 1) and 84 sera containing monoclonal IgG (Group 2) in assays in which we used either of two anti-IgG monoclonal antibodies (HG6 or HG8). Results compared well with those obtained with a nephelometric assay involving polyclonal antiserum, except for sera from four persons of Group 2 whose immunoglobulins were not detected by antibody HG6. HG6 bound IgG from these four sera in a solid-phase binding assay. HG6 and HG8 recognize epitopes on the Fab and Fc regions of IgG, respectively, and they do not compete for binding to the whole molecule. However, use of the two monoclonal antibodies combined failed to improve the sensitivity or range of the assay. We conclude that light-scattering assays of IgG can be validly performed with a single monoclonal antibody.
Y
Y
Maynard
M G
MG
Scott
M H
MH
Nahm
J H
JH
Ladenson
D000818:Animals; D000906:Antibodies / Q000276:immunology; D000911:Antibodies, Monoclonal; D006801:Humans; D007074:Immunoglobulin G / Q000032:analysis*; D051379:Mice; D009391:Nephelometry and Turbidimetry / Q000379:methods; D010710:Phosphates; D011092:Polyethylene Glycols / Q000494:pharmacology; D012965:Sodium Chloride; D013997:Time Factors
D000906:Antibodies; D000911:Antibodies, Monoclonal; D007074:Immunoglobulin G; D010710:Phosphates; D011092:Polyethylene Glycols; D012965:Sodium Chloride; C018279:sodium phosphate