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. 1971 Feb 28;133(3):424–441. doi: 10.1084/jem.133.3.424

RELATIONSHIPS BETWEEN RELATIVE BINDING AFFINITY AND ELECTROPHORETIC BEHAVIOR OF RABBIT ANTIBODIES TO STREPTOCOCCAL CARBOHYDRATES

K Eichmann 1, J Greenblatt 1
PMCID: PMC2138948  PMID: 4999997

Abstract

After repeated intravenous injections with Group C streptococcal vaccine, most rabbit antisera were shown to contain one or more IgG antibody components, as revealed by microzone electrophoresis. A procedure for the fractionation of multiple IgG antibody components from such streptococcal antisera is described. Separation is achieved on the basis of differences in relative binding affinities of the antibody components to immunoabsorbent columns. The evidence suggests that the electrophoretic mobility, and thus the net charge of an antibody, bears a reciprocal relationship to its binding affinity for the streptococcal Group C antigens. Furthermore, the relative binding affinity affords another means to assess the functional homogeneity of streptococcal antibodies. A possible relationship between light chain variable-region subclasses and binding affinities of streptococcal antibodies is discussed.

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Selected References

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

  1. Axén R., Porath J., Ernback S. Chemical coupling of peptides and proteins to polysaccharides by means of cyanogen halides. Nature. 1967 Jun 24;214(5095):1302–1304. doi: 10.1038/2141302a0. [DOI] [PubMed] [Google Scholar]
  2. Braun D. G., Eichmann K., Krause R. M. Rabbit antibodies to streptococcal carbohydrates. Influence of primary and secondary immunization and of possible genetic factors on the antibody response. J Exp Med. 1969 Apr 1;129(4):809–830. doi: 10.1084/jem.129.4.809. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Braun D. G., Krause R. M. The individual antigenic specificity of antibodies to streptococcal carbohydrates. J Exp Med. 1968 Nov 1;128(5):969–989. doi: 10.1084/jem.128.5.969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Cuatrecasas P., Wilchek M., Anfinsen C. B. Selective enzyme purification by affinity chromatography. Proc Natl Acad Sci U S A. 1968 Oct;61(2):636–643. doi: 10.1073/pnas.61.2.636. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Cutinelli C., Galdiero F. Ion-binding properties of the cell wall of Staphylococcus aureus. J Bacteriol. 1967 Jun;93(6):2022–2023. doi: 10.1128/jb.93.6.2022-2023.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Eichmann K., Braun D. G., Feizi T., Krause R. M. The emergence of antibodies with either identical or unrelated individual antigenic specificity during repeated immunizations with streptococcal vaccines. J Exp Med. 1970 Jun 1;131(6):1169–1189. doi: 10.1084/jem.131.6.1169. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Eichmann K., Lackland H., Hood L., Krause R. M. Induction of rabbit antibody with molecular uniformity after immunization with group C streptococci. J Exp Med. 1970 Jan 1;131(1):207–221. doi: 10.1084/jem.131.1.207. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Freedman M. H., Grossberg A. L., Pressman D. Participation of amino groups in the active sites of antibodies. Immunochemistry. 1968 Jul;5(4):367–381. doi: 10.1016/0019-2791(68)90132-8. [DOI] [PubMed] [Google Scholar]
  9. Henney C. S., Ishizaka K. A simplified procedure for the preparation of immunoglobulin-class-specific antisera. J Immunol. 1969 Jul;103(1):56–61. [PubMed] [Google Scholar]
  10. Hood L., Eichmann K., Lackland H., Krause R. M., Ohms J. J. Rabbit antibody light chains and gene evolution. Nature. 1970 Dec 12;228(5276):1040–1044. doi: 10.1038/2281040a0. [DOI] [PubMed] [Google Scholar]
  11. Jaton J. C., Waterfield M. D., Margolies M. N., Haber E. Isolation and characterization of structurally homogeneous antibodies from antipneumococcal sera. Proc Natl Acad Sci U S A. 1970 Jul;66(3):959–966. doi: 10.1073/pnas.66.3.959. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. KRAUSE R. M., McCARTY M. Studies on the chemical structure of the streptococcal cell wall. II. The composition of group C cell walls and chemical basis for serologic specificity of the carbohydrate moiety. J Exp Med. 1962 Jan 1;115:49–62. doi: 10.1084/jem.115.1.49. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Kindt T. J., Todd C. W., Eichmann K., Krause R. M. Allotype exclusion in uniform rabbit antibody to streptococcal carbohydrate. J Exp Med. 1970 Feb;131(2):343–354. doi: 10.1084/jem.131.2.343. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Kristiansen T., Sundberg L., Porath J. Studies on blood group substances. II. Coupling of blood group substane A to hydroxyl-containing matrices, including aminoethyl cellulose and agarose. Biochim Biophys Acta. 1969 Jun 17;184(1):93–98. [PubMed] [Google Scholar]
  15. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  16. Osterland C. K., Miller E. J., Karakawa W. W., Krause R. M. Characteristics of streptococcal group-specific antibody isolated from hyperimmune rabbits. J Exp Med. 1966 Apr 1;123(4):599–614. doi: 10.1084/jem.123.4.599. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Pincus J. H., Jaton J. C., Bloch K. J., Haber E. Antibodies to type III and type VIII pneumococcal polysaccharides: evidence for restricted structural heterogeneity in hyperimmunized rabbits. J Immunol. 1970 May;104(5):1143–1148. [PubMed] [Google Scholar]
  18. Pincus J. H., Jaton J. C., Bloch K. J., Haber E. Properties of structurally restricted antibody to type VIII pneumococcal polysaccharide. J Immunol. 1970 May;104(5):1149–1154. [PubMed] [Google Scholar]
  19. Porath J., Axen R., Ernback S. Chemical coupling of proteins to agarose. Nature. 1967 Sep 30;215(5109):1491–1492. doi: 10.1038/2151491a0. [DOI] [PubMed] [Google Scholar]
  20. Reisfeld R. A., Small P. A., Jr Electrophoretic heterogeneity of polypeptide chains of specific antibodies. Science. 1966 May 27;152(3726):1253–1255. doi: 10.1126/science.152.3726.1253. [DOI] [PubMed] [Google Scholar]
  21. Sela M., Mozes E. Dependence of the chemical nature of antibodies on the net electrical charge of antigens. Proc Natl Acad Sci U S A. 1966 Feb;55(2):445–452. doi: 10.1073/pnas.55.2.445. [DOI] [PMC free article] [PubMed] [Google Scholar]

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