Composition of IgA immune complexes precipitated with polyethylene glycol. A model for isolation and analysis of immune complexes

J Immunol Methods. 1987 Nov 5;103(2):239-45. doi: 10.1016/0022-1759(87)90295-x.

Abstract

Covalently cross-linked large and intermediate-sized IgA oligomers, prepared with IgA anti-dinitrophenyl (DNP) and bis-DNP-pimelic acid ester, were used to examine the ability of different concentrations (3.5%, 5% and 7%, w/v) of polyethylene glycol (PEG) to precipitate IgA immune complexes (IgA-IC). The size of the IgA-IC precipitated with PEG was determined by gradient polyacrylamide gel electrophoresis and quantitative autoradiography. The standard concentration of 3.5% PEG precipitated only a minor fraction (20%) of the IgA-IC. In contrast, 5% and 7% PEG precipitated 45% and 79% of the complexes, respectively. To test the influence of the antigen on the PEG assay. IgA-IC prepared with IgA anti-DNP and DNP conjugates of either bovine serum albumin or Ficoll were also used. Approximately 38% of these IgA-IC were precipitated with 3.5%, PEG. By comparison, the concentration of 5% and 7% PEG precipitated 60% and 76% of the IgA-IC, respectively. Distilled water rather than the standard borate-buffered saline was shown to be the optimal solvent for resolubilization of the PEG precipitates. Serum samples from 22 IgA nephropathy patients and 12 normal donors were tested with 3.5%, 5% and 7% PEG. Only the 7% PEG assay showed a significant difference between patients and controls (P less than 0.001) in the IgA levels of precipitates. Thus, the use of 7% PEG is recommended for the detection, isolation and analysis of large- and intermediate-sized IgA-IC.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Antigen-Antibody Complex / analysis*
  • Chemical Precipitation
  • Glomerulonephritis, IGA / immunology
  • Humans
  • Immunoglobulin A / analysis*
  • Molecular Weight
  • Polyethylene Glycols
  • Solubility
  • Structure-Activity Relationship

Substances

  • Antigen-Antibody Complex
  • Immunoglobulin A
  • Polyethylene Glycols