Abstract
Idiotypic antibodies were investigated quantitatively by a method of indirect precipitation, which utilizes labeled F(ab')2 fragments of specifically purified antibenzoate antibody from the donor, anti-antibody, and an antiglobulin reagent. The contribution of allotypic and hidden determinants to these reactions was excluded. Greater fractions of an idiotypic antibody population are precipitated by this method, as compared to direct precipitation, and in two instances large proportions of idiotypic antibodies were detected in populations which failed to form precipitates by double diffusion in agar gel. The greater sensitivity of the indirect method was attributed to its capacity to detect molecules bearing a small number of antigenic determinants. Extensive studies of cross-reactions, carried out by an inhibition technique, failed to reveal any strong reactions of anti-idiotypic antibodies with heterologous antibenzoate antibody preparations, heterologous sera, or IgG, although a few weak cross-reactions were noted. One definite cross-reaction was observed by a direct binding measurement with heterologous antiserum. Antisera prepared in more than one recipient against a single donor preparation reacted with identical or overlapping subpopulations of the donor molecules. Instances in which two recipient antisera reacted with different proportions of the molecules of a single donor provided evidence for the existence of more than one idiotypic antibody population in the antibenzoate antibody of an individual rabbit.
Full Text
The Full Text of this article is available as a PDF (926.6 KB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- 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]
- Cohn M., Notani G., Rice S. A. Characterization of the antibody to the C-carbohydrate produced by a transplantable mouse plasmacytoma. Immunochemistry. 1969 Jan;6(1):111–123. doi: 10.1016/0019-2791(69)90183-9. [DOI] [PubMed] [Google Scholar]
- Daugharty H., Hopper J. E., MacDonald A. B., Nisonoff A. Quantitative investigations of idiotypic antibodies. I. Analysis of precipitating antibody populations. J Exp Med. 1969 Nov 1;130(5):1047–1062. doi: 10.1084/jem.130.5.1047. [DOI] [PMC free article] [PubMed] [Google Scholar]
- GILMAN A. M., NISONOFF A., DRAY S. SYMMETRICAL DISTRIBUTION OF GENETIC MARKERS IN INDIVIDUAL RABBIT GAMMA-GLOBULIN MOLECULES. Immunochemistry. 1964 Jun;1:109–120. doi: 10.1016/0019-2791(64)90075-8. [DOI] [PubMed] [Google Scholar]
- GREY H. M., MANNIK M., KUNKEL H. G. INDIVIDUAL ANTIGENIC SPECIFICITY OF MYELOMA PROTEINS. CHARACTERISTICS AND LOCALIZATION TO SUBUNITS. J Exp Med. 1965 Apr 1;121:561–575. doi: 10.1084/jem.121.4.561. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hurez D., Meshaka G., Mihaesco C., Seligmann M. The inhibition by normal gamma-G-globulins of antibodies specific for individual gamma-G-myeloma proteins. J Immunol. 1968 Jan;100(1):69–79. [PubMed] [Google Scholar]
- Kelus A. S., Gell P. G. Immunological analysis of rabbit anti-antibody systems. J Exp Med. 1968 Jan 1;127(1):215–234. doi: 10.1084/jem.127.1.215. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kunkel H. G., Mannik M., Williams R. C. Individual Antigenic Specificity of Isolated Antibodies. Science. 1963 Jun 14;140(3572):1218–1219. doi: 10.1126/science.140.3572.1218. [DOI] [PubMed] [Google Scholar]
- NISONOFF A., WISSLER F. C., LIPMAN L. N., WOERNLEY D. L. Separation of univalent fragments from the bivalent rabbit antibody molecule by reduction of disulfide bonds. Arch Biochem Biophys. 1960 Aug;89:230–244. doi: 10.1016/0003-9861(60)90049-7. [DOI] [PubMed] [Google Scholar]
- Oudin J. The genetic control of immunoglobulin synthesis. Proc R Soc Lond B Biol Sci. 1966 Nov 22;166(1003):207–221. doi: 10.1098/rspb.1966.0094. [DOI] [PubMed] [Google Scholar]
- PORTER R. R. The hydrolysis of rabbit y-globulin and antibodies with crystalline papain. Biochem J. 1959 Sep;73:119–126. doi: 10.1042/bj0730119. [DOI] [PMC free article] [PubMed] [Google Scholar]
- SKOM J. H., TALMAGE D. W. Nonprecipitating insulin antibodies. J Clin Invest. 1958 Jun;37(6):783–786. doi: 10.1172/JCI103664. [DOI] [PMC free article] [PubMed] [Google Scholar]
- UTIGER R. D., PARKER M. L., DAUGHADAY W. H. Studies on human growth hormone. I. A radio-immunoassay for human growth hormone. J Clin Invest. 1962 Feb;41:254–261. doi: 10.1172/JCI104478. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Willims R. C., Jr, Kunkel H. G., Capra J. D. Antigenic specificities related to the cold agglutinin activity of gamma M globulins. Science. 1968 Jul 26;161(3839):379–381. doi: 10.1126/science.161.3839.379. [DOI] [PubMed] [Google Scholar]