Abstract
We describe a simple and rapid method to dissociate antigen-antibody complexes to allow antigen detection by radioimmunoassay. Immunoglobulins and immune complexes were precipitated from serum in 50%-saturated ammonium sulfate. The precipitate was dissolved in distilled water and heated at 90 degrees C for 15 min to free the antigen and denature the antibody irreversibly. With this treatment, the sensitivity for detecting staphylococcal antigen added to human serum improved greater than or equal to 16-fold and was similar to the sensitivity in buffer. After intravenous injection of antigen into a rabbit with high levels of staphylococcal antibodies, antigen was detected only after thermodissociation. Also, in 4 of 26 patients with Staphylococcus aureus bacteremia, circulating antigen was detected after thermodissociation. Thermodissociation of immune complexes is a simple, rapid method and should be useful for detecting other heat-stable antigens.
Full text
PDF






Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Bayer A. S., Theofilopoulos A. N., Dixon F. J., Guze L. B. Circulating immune complexes in experimental streptococcal endocarditis: a monitor of therapeutic efficacy. J Infect Dis. 1979 Jan;139(1):1–8. doi: 10.1093/infdis/139.1.1. [DOI] [PubMed] [Google Scholar]
- Bayer A. S., Theofilopoulos A. N., Eisenberg R., Dixon F. J., Guze L. B. Circulating immune complexes in infective endocarditis. N Engl J Med. 1976 Dec 30;295(27):1500–1505. doi: 10.1056/NEJM197612302952703. [DOI] [PubMed] [Google Scholar]
- Bayer A. S., Theofilopoulos A. N., Tillman D. B., Dixon F. J., Guze L. B. Use of circulating immune complex levels in the serodifferentiation of endocarditic and nonendocarditic septicemias. Am J Med. 1979 Jan;66(1):58–62. doi: 10.1016/0002-9343(79)90482-0. [DOI] [PubMed] [Google Scholar]
- Carella G., Digeon M., Feldmann G., Jungers P., Drouet J., Bach J. F. Detection of hepatitis B antigen in circulating immune complexes in acute and chronic hepatitis. Scand J Immunol. 1977;6(12):1297–1304. doi: 10.1111/j.1365-3083.1977.tb00369.x. [DOI] [PubMed] [Google Scholar]
- Coonrod J. D., Leach R. P. Immunoelectrophoresis for detection of polysaccharides in immune complexes. J Clin Microbiol. 1978 Aug;8(2):257–259. doi: 10.1128/jcm.8.2.257-259.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Coonrod J. D., Rylko-Bauer Latex agglutination in the diagnosis of pneumococcal infection. J Clin Microbiol. 1976 Aug;4(2):168–174. doi: 10.1128/jcm.4.2.168-174.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gocke D. J., Howe C. Rapid detection of Australia antigen by counterimmunoelectrophoresis. J Immunol. 1970 Apr;104(4):1031–1034. [PubMed] [Google Scholar]
- Kohler R. B., Wheat L. J., White A. Rapid diagnosis of Pseudomonas aeruginosa urinary tract infections by radioimmunoassay. J Clin Microbiol. 1979 Feb;9(2):253–258. doi: 10.1128/jcm.9.2.253-258.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lampe R. M., Chottipitayasunondh T., Sunakorn P. Detection of bacterial antigen in pleural fluid by counterimmunoelectrophoresis. J Pediatr. 1976 Apr;88(4 Pt 1):557–560. doi: 10.1016/s0022-3476(76)80005-4. [DOI] [PubMed] [Google Scholar]
- Leach R. P., Coonrod J. D. Detection of penumococcal antigens in the sputum in pneumococcal pneumonia. Am Rev Respir Dis. 1977 Nov;116(5):847–851. doi: 10.1164/arrd.1977.116.5.847. [DOI] [PubMed] [Google Scholar]
- Madaliński K., Sztachelska-Budkowska A., Brzosko W. J. DEAE-cellulose chromatography: a method for dissociation of soluble immune complexes of hepatitis B antigen. J Infect Dis. 1974 Apr;129(4):371–375. doi: 10.1093/infdis/129.4.371. [DOI] [PubMed] [Google Scholar]
- Newman R. B., Stevens R. W., Gaafar H. A. Latex agglutination test for the diagnosis of Haemophilus influenzae meningitis. J Lab Clin Med. 1970 Jul;76(1):107–113. [PubMed] [Google Scholar]
- O'Reilly R. J., Anderson P., Ingram D. L., Peter G., Smith D. H. Circulating polyribophosphate in Hemophilus influenzae, type b meningitis. Correlation with clinical course and antibody response. J Clin Invest. 1975 Oct;56(4):1012–1022. doi: 10.1172/JCI108148. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Overby L. R., Miller J. P., Smith I. D., Decker R. H., Ling C. M. Radioimmunoassay of hepatitis B virus-associated (Australia) antigen employing 125 I-antibody. Vox Sang. 1973;24(Suppl):102–113. doi: 10.1111/j.1423-0410.1973.tb03523.x. [DOI] [PubMed] [Google Scholar]
- Peterson P. K., Wilkinson B. J., Kim Y., Schmeling D., Douglas S. D., Quie P. G., Verhoef J. The key role of peptidoglycan in the opsonization of Staphylococcus aureus. J Clin Invest. 1978 Mar;61(3):597–609. doi: 10.1172/JCI108971. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Popova O. Y., Kositskaya L. S. Dissociation of immune complexes and inactivation of bound antibodies by reducing agents. Immunochemistry. 1977 Aug;14(8):633–635. doi: 10.1016/0019-2791(77)90161-6. [DOI] [PubMed] [Google Scholar]
- Raam S., Chang C. H., Fishman W. H. Freezing and thawing as a means of dissociating antigen bound to immobilized antibody. Immunochemistry. 1975 Sep;12(9):723–726. doi: 10.1016/0019-2791(75)90221-9. [DOI] [PubMed] [Google Scholar]
- Ribner B., Keusch G. T., Robbins J. B. Letter: Staphylococcus aureus antigen in cerebrospinal fluid cross-reactive with Haemophilus influenzae type b antiserum. Ann Intern Med. 1975 Sep;83(3):370–371. doi: 10.7326/0003-4819-83-3-370. [DOI] [PubMed] [Google Scholar]
- Rolleri E., Malvano R., Gandolfi C., Rosa U. Effect of irreversible adsorbents on the dissociation of the antigen-antibody complexes. Horm Metab Res. 1974 Jan;6(1):57–61. doi: 10.1055/s-0028-1093885. [DOI] [PubMed] [Google Scholar]
- Sugerman D., Hart H. E. Thermal dissociation of antigen-antibody-antigen-like systems. Bull Math Biol. 1973 Feb-Apr;35(1):219–235. doi: 10.1007/BF02558808. [DOI] [PubMed] [Google Scholar]
- Tabbarah Z. A., Kohler R. B., Wheat L. J., Griep J. A., White A. Inhibitory effect of heat-labile serum factors on detection of staphylococcal, pseudomonas, and hepatitis B surface antigens by solid-phase radioimmunoassay. J Infect Dis. 1979 Nov;140(5):822–825. doi: 10.1093/infdis/140.5.822. [DOI] [PubMed] [Google Scholar]
- 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]
- WILLIAMS R. C., Jr, KUNKEL H. G. Rheumatoid factor, complement, and conglutinin aberrations in patients with subacute bacterial endocarditis. J Clin Invest. 1962 Mar;41:666–675. doi: 10.1172/JCI104523. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wheat L. J., Humphreys D. W., White A. Opsonization of staphylococci by normal human sera: the role of antibody and heat-labile factors. J Lab Clin Med. 1974 Jan;83(1):73–78. [PubMed] [Google Scholar]
- Wheat L. J., Kohler R. B., White A. Solid-phase radioimmunoassay for detection of staphylococcal antigen in serum of rabbits with endocraditis due to Staphylococcus aureus. J Infect Dis. 1978 Aug;138(2):174–180. doi: 10.1093/infdis/138.2.174. [DOI] [PubMed] [Google Scholar]
- Wheat L. J., Kohler R. B., White A. Solid-phase radioimmunoassay for immunoglobulin G Staphylococcus aureus antibody in serious staphylococcal infection. Ann Intern Med. 1978 Oct;89(4):467–472. doi: 10.7326/0003-4819-89-4-467. [DOI] [PubMed] [Google Scholar]
