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Measurement of Antibody Affinity, Concentration and Isotype to Evaluate Antigens, Adjuvants, and Immunization Regimens in Vaccine Research

  • Brian W. Hughes
  • John S. Kenney
  • Anthony C. Allison
Part of the NATO ASI Series book series (NSSA, volume 179)

Abstract

The recent explosion in vaccine research has brought into focus the need for better methods of characterizing the serum antibodies produced in animals immunized with various adjuvants and antigens. The typical end-point ELISAs used to characterize sera fail to differentiate between antibody affinity and concentration and cannot quantitate antibody isotypes. Antibody affinity and concentration when measured directly can serve as invariant standards to judge the quality of immunization and are not subject to arbitrary interpretation that is applied to end-point titer ELISAs. Furthermore, antibody affinity determines the effectiveness of antibody in j protection (reviewed in Steward and Steensgard, 1983). Antibody isotypes, which are often neglected in evaluating vaccines, have also been shown to have a marked effect on the generation of immunoprophylaxis (Wechsler and Kongshavn, 1986). We therefore suggest that measurements of antibody affinity, concentration, and isotype should be included in the evaluation of the efficacy of a vaccine. To measure these characteristics, we have developed a convenient solution-phase RIA for antibody affinity and concentration and an ELISA for antibody isotypes.

Keywords

Human Serum Albumin Incubation Buffer Antibody Isotype Antibody Affinity Vaccine Research 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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References

  1. Apple, R., Knauper, B., Pesce, A. and Michael, G., 1984, Shared antigenic determinants of native and denatured bovine serum albumin are recognized by both B-and T-cells, Mol. Immunol., 21: 901.PubMedCrossRefGoogle Scholar
  2. Gani, M., Coley, J. and Porter, P., 1987, Epitope masking and immunodominance — complications in the selection of monoclonal antibodies against HCG, Hybridoma, 6: 637.PubMedCrossRefGoogle Scholar
  3. Kenney, J.S., Hughes, B.H., Masada, M.P. and Allison, A.C., 1989a, Influence of adjuvants on the quantity, affinity, isotype, and epitope specificity of murine antibodies, J. Immunol. Methods, in press.Google Scholar
  4. Kenney, J.S., Hughes, B.H. and Allison, A.C., 1989b, Determination of antibody affinity and concentration by solution-phase micro-radioimmunoassay, in: “Focus on Laboratory Methods in Immunology”, H. Zola, ed., CRC Press, Boca Raton.Google Scholar
  5. Muller, R., 1980, Calculation of average antibody affinity in anti-hapten sera from data obtained by competitive radioimmunoassay, J. Immunol. Methods, 34: 345.PubMedCrossRefGoogle Scholar
  6. Steward, M.W. and Petty, R.E., 1972, The use of ammonium sulphate globulin precipitation for determination of affinity of anti-protein antibodies in mouse serum, Immunology, 22: 747.PubMedGoogle Scholar
  7. Steward, M.W. and Petty, R.E., 1976, Evidence for the genetic control of antibody affinity from breeding studies with inbred mouse strains producing high and low affinity antibody, Immunology, 30: 789.PubMedGoogle Scholar
  8. Steward, M.W. and Steensgaard, J., 1983, Antibody affinity: thermodynamic aspects and biological significance, CRC Press, Boca Raton.Google Scholar
  9. Wechsler, D.S. and Kongshavn, P.A.L., 1986, Heat-labile IgG2a antibodies affect cure of Trypanosoma musculi infection in C57BL/6 mice, J. Immunol., 137: 2968.PubMedGoogle Scholar
  10. Wolberg, G., Liu, C.T. and Adler, F.L., 1970, Passive hemagglutination. II. Titration of antibody against determinants unique for aggregated denatured bovine serum albumin and further studies on gelatin, J. Immunol., 105: 797.PubMedGoogle Scholar

Copyright information

© Plenum Press, New York 1989

Authors and Affiliations

  • Brian W. Hughes
    • 1
  • John S. Kenney
    • 1
  • Anthony C. Allison
    • 1
  1. 1.Department of ImmunologySyntex ResearchPalo AltoUSA

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