Skip to main content
PMC home page
Clinical and Experimental Immunology logoLink to Clinical and Experimental Immunology
. 1983 Jan;51(1):8–16.

Separation of two molecular species of the Sm antigen by affinity chromatography with murine monoclonal and human anti-nuclear autoantibodies.

R R Buchanan, P J Venables, A Morgan, N A Staines, P R Smith, R N Maini
PMCID: PMC1536750  PMID: 6403266

Abstract

Nuclear ribonuclear protein (nRNP) and Sm were extracted from a 30-60% ammonium sulphate fraction of rabbit thymus extract by affinity chromatography. Immunoadsorbent columns were prepared from IgG extracted from an anti-nRNP serum, an anti-Sm serum and two monoclonal autoantibodies derived from a MRL/lpr mouse. All four immunoadsorbents isolated both nRNP and Sm antigens indicating that they exist as the 'nRNP/Sm complex' A species of Sm which did not bind to the anti-nRNP column and which was subsequently purified with the anti-Sm column was termed 'free Sm'. The Sm in the complex and free Sm were immunologically identical on immunodiffusion and gave similar polypeptide bands on polyacrylamide gel electrophoresis. However, they differed in that complexed Sm was sensitive to heat and RNAase. These studies provide direct evidence of a physical association between the nRNP and Sm antigens and indicate an additional molecular species of Sm whose resistance to RNAase may be due to different associated RNA species.

Full text

PDF

Images in this article

11Fig. 1
on p.11
12Fig. 3
on p.12
13Fig. 4
on p.13

Selected References

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

  1. Galfre G., Howe S. C., Milstein C., Butcher G. W., Howard J. C. Antibodies to major histocompatibility antigens produced by hybrid cell lines. Nature. 1977 Apr 7;266(5602):550–552. doi: 10.1038/266550a0. [DOI] [PubMed] [Google Scholar]
  2. Kearney J. F., Radbruch A., Liesegang B., Rajewsky K. A new mouse myeloma cell line that has lost immunoglobulin expression but permits the construction of antibody-secreting hybrid cell lines. J Immunol. 1979 Oct;123(4):1548–1550. [PubMed] [Google Scholar]
  3. Lerner M. R., Steitz J. A. Antibodies to small nuclear RNAs complexed with proteins are produced by patients with systemic lupus erythematosus. Proc Natl Acad Sci U S A. 1979 Nov;76(11):5495–5499. doi: 10.1073/pnas.76.11.5495. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Mattioli M., Reichlin M. Physical association of two nuclear antigens and mutual occurrence of their antibodies: the relationship of the SM and RNAprotein (MO) systems in SLE sera. J Immunol. 1973 May;110(5):1318–1324. [PubMed] [Google Scholar]
  5. Notman D. D., Kurata N., Tan E. M. Profiles of antinuclear antibodies in systemic rheumatic diseases. Ann Intern Med. 1975 Oct;83(4):464–469. doi: 10.7326/0003-4819-83-4-464. [DOI] [PubMed] [Google Scholar]
  6. Peltier A. P., Aussel C., Haim T., Cyna L. Ribonucleoprotein (RNP) soluble nuclear antigen: demonstration of its reaction with serum antibodies by counter-immunoelectrophoresis and further partial characterization. J Immunol Methods. 1977;16(2):153–163. doi: 10.1016/0022-1759(77)90050-3. [DOI] [PubMed] [Google Scholar]
  7. Staines N. A., Hardingham T., Smith M., Henderson B. Collagen-induced arthritis in the rat: modification of immune and arthritic responses by free collagen and immune anti-collagen antiserum. Immunology. 1981 Dec;44(4):737–744. [PMC free article] [PubMed] [Google Scholar]
  8. Takano M., Agris P. F., Sharp G. C. Purification and biochemical characterization of nuclear ribonucleoprotein antigen using purified antibody from serum of a patient with mixed connective tissue disease. J Clin Invest. 1980 Jun;65(6):1449–1456. doi: 10.1172/JCI109809. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Tan E. M., Kunkel H. G. Characteristics of a soluble nuclear antigen precipitating with sera of patients with systemic lupus erythematosus. J Immunol. 1966 Mar;96(3):464–471. [PubMed] [Google Scholar]
  10. Venables P. J., Erhardt C. C., Maini R. N. Antibodies to extractable nuclear antigens in rheumatoid arthritis: relationship to vasculitis and circulating immune complexes. Clin Exp Immunol. 1980 Jan;39(1):146–153. [PMC free article] [PubMed] [Google Scholar]
  11. Waelti E. R., Hess M. W. Nuclear antigens: characterization of the acidic nuclear ribonucleoprotein-Sm-antigen complex by analytical and preparative isoelectric focusing. J Immunol Methods. 1980;32(2):177–184. doi: 10.1016/0022-1759(80)90071-x. [DOI] [PubMed] [Google Scholar]
  12. White P. J., Hoch S. O. Definition of the antigenic polypeptides in the Sm and RNP ribonucleoprotein complexes. Biochem Biophys Res Commun. 1981 Sep 16;102(1):365–371. doi: 10.1016/0006-291x(81)91530-8. [DOI] [PubMed] [Google Scholar]

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

RESOURCES