Development of an immunoassay for the detection of minute amounts of IgG-coated erythrocytes in whole blood and its application for the assessment of Fc-mediated clearance of anti-D-coated erythrocytes in vivo

Vox Sang. 1989;57(3):188-92. doi: 10.1111/j.1423-0410.1989.tb00821.x.

Abstract

A rapid and sensitive immunoassay for the detection of minute quantities of IgG-coated erythrocytes in whole blood was developed. Washed red blood cells were incubated in two steps with anti-human IgG antiserum followed by 125I-labelled protein A. The assay was able to detect amounts of sensitized erythrocytes as small as 0.5 ml of packed erythrocytes in a total blood volume of 5 liters and hematocrit 40%. A linear relation between increasing amounts of IgG-coated red cells in whole blood and the binding of 125I-labelled protein A was obtained. We applied the technique on the assessment of the removal of IgG anti-D-coated erythrocytes from the circulation of test individuals. T1/2 for the elimination of approximately 4 ml packed red cells sensitized with 62 micrograms of anti-D in 14 normal subjects was 20 +/- 5 min (mean +/- SEM). A splenectomized person did not clear the injected cells from the circulation during the test period of 70 min. If a standard curve was constructed the total blood volume in the test subjects could be calculated. This value correlated well (r = 0.99) with the blood volume calculated from the height and weight of the test individuals.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Erythrocytes / immunology*
  • Erythrocytes / metabolism
  • Female
  • Humans
  • Immunoassay / methods
  • Immunoglobulin Fc Fragments / physiology
  • Immunoglobulin G / analysis*
  • Immunoglobulin G / metabolism
  • Immunoglobulins / analysis*
  • Immunoglobulins / metabolism
  • Isoantibodies / analysis*
  • Male
  • Middle Aged
  • Rho(D) Immune Globulin
  • Splenectomy

Substances

  • Immunoglobulin Fc Fragments
  • Immunoglobulin G
  • Immunoglobulins
  • Isoantibodies
  • Rho(D) Immune Globulin