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. 1960 Nov 30;112(6):1195–1210. doi: 10.1084/jem.112.6.1195

THE PREPARATION AND SOME PROPERTIES OF PURIFIED ANTIBODY SPECIFIC FOR THE 2,4-DINITROPHENYL GROUP

Fuad S Farah 1, Milton Kern 1, Herman N Eisen 1
PMCID: PMC2137313  PMID: 13698150

Abstract

Injection of rabbits with Freund's complete adjuvant containing bovine γ-globulin substituted to near saturation with dinitrophenyl groups leads to the production of large amounts of antibodies specific for the 2,4-dinitrophenyl substituent. These antibodies can be isolated from sera in satisfactory yield (40 per cent) and in a high state of purity (90 per cent specifically precipitable). Hapten dissolution, which was used to screen haptens for their potential usefulness as specific reagents in the purification procedure, provides a means for evaluating, in relative terms, antibody-hapten affinities (in competition with antigen).

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Selected References

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

  1. DE WECK A. L., EISEN H. N. Some immunochemical properties of penicillenic acid. An antigenic determinant derived from penicillin. J Exp Med. 1960 Dec 1;112:1227–1247. doi: 10.1084/jem.112.6.1227. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. EISEN H. N., CARSTEN M. E., BELMAN S. Studies of hypersensitivity to low molecular weight substances. III. The 2,4-dinitrophenyl group as a determinant in the preciptin reaction. J Immunol. 1954 Nov;73(5):296–308. [PubMed] [Google Scholar]
  3. EISEN H. N., KERN M., NEWTON W. T., HELMREICH E. A study of the distribution of 2,4-dinitrobenzene sensitizers between isolated lymph node cells and extracellular medium in relation to induction of contact skin sensitivity. J Exp Med. 1959 Aug 1;110(2):187–206. doi: 10.1084/jem.110.2.187. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. EISEN H. N., TABACHNICK M. Elicitation of allergic contact dermatitis in the guinea pig; the distribution of bound dinitrobenzene groups within the skin and quantitative determination of the extent of combination of 2,4-dinitro-chlorobenzene with epidermal protein in vivo. J Exp Med. 1958 Dec 1;108(6):773–796. doi: 10.1084/jem.108.6.773. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. FARAH F. S., KERN M., EISEN H. N. Specific inhibition of wheal-and-erythema responses with univalent haptens and univalent antibody fragments. J Exp Med. 1960 Dec 1;112:1211–1226. doi: 10.1084/jem.112.6.1211. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. GITLIN D. Use of ultraviolet absorption spectroscopy in the quantitative precipitin reaction. J Immunol. 1949 Aug;62(4):437–451. [PubMed] [Google Scholar]
  7. Heidelberger M., Kendall F. E. QUANTITATIVE STUDIES ON ANTIBODY PURIFICATION : I. THE DISSOCIATION OF PRECIPITATES FORMED BY PNEUMOCOCCUS SPECIFIC POLYSACCHARIDES AND HOMOLOGOUS ANTIBODIES. J Exp Med. 1936 Jul 31;64(2):161–172. doi: 10.1084/jem.64.2.161. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. KABAT E. A. Heterogeneity in extent of the combining regions of human antidextran. J Immunol. 1956 Dec;77(6):377–385. [PubMed] [Google Scholar]
  9. KABAT E. A., MCDUFFIE F. C. A comparative study of methods used for analysis of specific precipitates in quantitative immunochemistry. J Immunol. 1956 Sep;77(3):193–197. [PubMed] [Google Scholar]
  10. KARUSH F., MARKS R. The preparation and properties of purified anti-hapten antibody. J Immunol. 1957 Apr;78(4):296–303. [PubMed] [Google Scholar]
  11. Kern M., Helmreich E., Eisen H. N. A DEMONSTRATION OF ANTIBODY ACTIVITY ON MICROSOMES. Proc Natl Acad Sci U S A. 1959 Jun;45(6):862–867. doi: 10.1073/pnas.45.6.862. [DOI] [PMC free article] [PubMed] [Google Scholar]

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