Skip to main content
NCBI home page
Clinical and Experimental Immunology logoLink to Clinical and Experimental Immunology
. 1981 Oct;46(1):44–53.

Detection of specific antigen within circulating immune complexes: validation of the assay and its application to food antigen-antibody complexes formed in healthy and food-allergic subjects.

R Paganelli, R J Levinsky, D J Atherton
PMCID: PMC1536342  PMID: 6896019

Abstract

A simple two-step method for the detection of specific antigen within immune complexes is described. The immune complexes are precipitated from serum by polyethylene glycol, dissociated by incubation in acid pH buffer and adsorbed onto the surface of polystyrene tubes. The antigen is detected by the binding of a radiolabelled affinity-purified specific antibody. The assay can detect the antigen within both antigen- and antibody-excess immune complexes of any immunoglobulin class, and can also allow semiquantitative comparison of different samples. Immune complexes containing food protein antigens after eating have been found in the serum of both normal subjects and atopic patients; the latter group showed higher mean levels of antigen-specific immune complexes. The method can be adopted for large-scale screening of clinical samples for suspected antigens if suitable antisera are available.

Full text

PDF
44

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Casali P., Lambert P. H. Purification of soluble immune complexes from serum using polymethylmetacrylate beads coated with conglutinin or C1q. Application to the analysis of the components of in vitro formed immune complexes and of immune complexes occurring in vivo during leishmaniasis. Clin Exp Immunol. 1979 Aug;37(2):295–309. [PMC free article] [PubMed] [Google Scholar]
  2. Chenais F., Virella G., Patrick C. C., Fudenberg H. H. Isolation of soluble immune complexes by affinity chromatography using staphylococcal protein A--Sepharose as substrate. J Immunol Methods. 1977;18(1-2):183–192. doi: 10.1016/0022-1759(77)90169-7. [DOI] [PubMed] [Google Scholar]
  3. Chia D., Barnett E. V., Yamagata J., Knutson D., Restivo C., Furst D. Quantitation and characterization of soluble immune complexes precipitated from sera by polyethylene glycol (PEG). Clin Exp Immunol. 1979 Sep;37(3):399–407. [PMC free article] [PubMed] [Google Scholar]
  4. Creighton W. D., Lambert P. H., Miescher P. A. Detection of antibodies and soluble antigen-antibody complexes by precipitation with polyethylene glycol. J Immunol. 1973 Oct;111(4):1219–1227. [PubMed] [Google Scholar]
  5. Dambuyant C., Burton-Kee J., Mowbray J. F. Demonstration of two disease specific antigens in circulating immune complexes. Clin Exp Immunol. 1979 Sep;37(3):424–431. [PMC free article] [PubMed] [Google Scholar]
  6. Digeon M., Laver M., Riza J., Bach J. F. Detection of circulating immune complexes in human sera by simplified assays with polyethylene glycol. J Immunol Methods. 1977;16(2):165–183. doi: 10.1016/0022-1759(77)90051-5. [DOI] [PubMed] [Google Scholar]
  7. Inganäs M., Johansson S. G., Dannaeus A. A method for estimation of circulating immune complexes after oral challenge with ovalbumin. Clin Allergy. 1980 May;10(3):293–302. doi: 10.1111/j.1365-2222.1980.tb02110.x. [DOI] [PubMed] [Google Scholar]
  8. Jayarao K. S., Faulk W. P., Karam J. H., Grodsky G. M., Forsham P. H. Measurement of immune complexes in insulin-treated diabetics. J Immunol Methods. 1973 Dec;3(4):337–345. doi: 10.1016/0022-1759(73)90035-5. [DOI] [PubMed] [Google Scholar]
  9. Mancini G., Carbonara A. O., Heremans J. F. Immunochemical quantitation of antigens by single radial immunodiffusion. Immunochemistry. 1965 Sep;2(3):235–254. doi: 10.1016/0019-2791(65)90004-2. [DOI] [PubMed] [Google Scholar]
  10. Masson P. L., Delire M., Cambiaso C. L. Circulating immune complexes in normal human pregnancy. Nature. 1977 Apr 7;266(5602):542–543. doi: 10.1038/266542a0. [DOI] [PubMed] [Google Scholar]
  11. May C. D., Remigio L., Feldman J., Bock S. A., Carr R. I. A study of serum antibodies to isolated milk proteins and ovalbumin in infants and children. Clin Allergy. 1977 Nov;7(6):583–595. doi: 10.1111/j.1365-2222.1977.tb01489.x. [DOI] [PubMed] [Google Scholar]
  12. Natali P. G., Walker L., Pellegrino M. A. Isolation of soluble immune complexes from serum using protein A bearing Staphylococcus aureus bacteria: separation of the antigen from immune complex and production of antisera. Clin Immunol Immunopathol. 1980 Jan;15(1):76–87. doi: 10.1016/0090-1229(80)90022-7. [DOI] [PubMed] [Google Scholar]
  13. PETERSON R. D., GOOD R. A. Antibodies to cow's milk proteins. Their presence and significance. Pediatrics. 1963 Feb;31:209–221. [PubMed] [Google Scholar]
  14. Paganelli R., Levinsky R. J., Brostoff J., Wraith D. G. Immune complexes containing food proteins in normal and atopic subjects after oral challenge and effect of sodium cromoglycate on antigen absorption. Lancet. 1979 Jun 16;1(8129):1270–1272. doi: 10.1016/s0140-6736(79)92230-x. [DOI] [PubMed] [Google Scholar]
  15. Paganelli R., Levinsky R. J. Solid phase radioimmunoassay for detection of circulating food protein antigens in human serum. J Immunol Methods. 1980;37(3-4):333–341. doi: 10.1016/0022-1759(80)90319-1. [DOI] [PubMed] [Google Scholar]
  16. Rubin R. L., Hardtke M. A., Carr R. I. The effect of high antigen density on solid-phase radioimmunoassays for antibody regardless of immunoglobulin class. J Immunol Methods. 1980;33(3):277–292. doi: 10.1016/0022-1759(80)90215-x. [DOI] [PubMed] [Google Scholar]
  17. Svehag S. E., Burger D. Isolation of C1q-binding immune complexes by affinity chromatography and desorption with a diaminoalkyl compound. Acta Pathol Microbiol Scand C. 1976 Feb;84(1):45–52. doi: 10.1111/j.1699-0463.1976.tb03598.x. [DOI] [PubMed] [Google Scholar]
  18. Theofilopoulos A. N., Eisenberg R. A., Dixon F. J. Isolation of circulating immune complexes using Raji cells. Separation of antigens from immune complexes and production of antiserum. J Clin Invest. 1978 Jun;61(6):1570–1581. doi: 10.1172/JCI109077. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Walker W. A., Isselbacher K. J. Uptake and transport of macromolecules by the intestine. Possible role in clinical disorders. Gastroenterology. 1974 Sep;67(3):531–550. [PubMed] [Google Scholar]

Articles from Clinical and Experimental Immunology are provided here courtesy of British Society for Immunology

RESOURCES