Monoclonal dinitrophenyl-specific murine IgE antibody: Preparation, isolation, and characterization

Fu-Tong Liu, J. W. Bohn, E. L. Ferry, H. Yamamoto, C. A. Molinaro, L. A. Sherman, N. R. Klinman, D. H. Katz

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Abstract

A murine hybridoma secreting monoclonal IgE antibodies of anti-2,4-dinitrophenyl (DNP) specificity was generated by fusion of SP2/0 tumor cells and spleen cells from DNP-Ascaris-hyperimmunized mice. Hybridomas secreting anti-DNP antibodies of other heavy chain classes, i.e., μ, γ 1 and γ(2b), were also obtained from the same fusion experiment. Large quantities of IgE antibodies were obtained from ascites of mice in which the IgE-secreting hybridoma was propagated in vivo. The IgE antibodies were isolated by precipitation with ammonium sulfate followed by affinity chromatography on DNP-bovine serum albumin (BSA)-Sepharose-4B and further purified by ion-exchange chromatography on DEAE-cellulose and gel filtration on Sephadex G-200. The isolated IgE has an approximate m.w. of 184,000, a total carbohydrate content of 13.3%, and its amino acid composition has been determined. The antibody has an association constant with DNP-lysine of 1.4 x 10 8 M -1 at 25°C and 10 7 7 M -1 at 37°C. Rabbit and goat antibodies against the hybridoma IgE were prepared and the antisera were made specific for IgE by adsorption on normal mouse serum protein-Sepharose-4B. Solid phase radioimmunoassays for measuring murine antigen-specific and total IgE were developed and were shown to have high specificity and sensitivity. Finally, the isolated hybridoma IgE can mediate antigen (DNP-BSA)-induced release of mediator (serotonin) from rat basophilic leukemia cells.

Original languageEnglish (US)
Pages (from-to)2728-2737
Number of pages10
JournalJournal of Immunology
Volume124
Issue number6
StatePublished - 1980

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Immunoglobulin E
Hybridomas
Antibodies
Sepharose
Ascaris
Antigens
DEAE-Cellulose
Ion Exchange Chromatography
Ammonium Sulfate
Affinity Chromatography
Ascites
Goats
Adsorption
Radioimmunoassay
Gel Chromatography
Blood Proteins
Immune Sera
Anti-Idiotypic Antibodies
Serotonin
Leukemia

ASJC Scopus subject areas

  • Immunology

Cite this

Liu, F-T., Bohn, J. W., Ferry, E. L., Yamamoto, H., Molinaro, C. A., Sherman, L. A., ... Katz, D. H. (1980). Monoclonal dinitrophenyl-specific murine IgE antibody: Preparation, isolation, and characterization. Journal of Immunology, 124(6), 2728-2737.

Monoclonal dinitrophenyl-specific murine IgE antibody : Preparation, isolation, and characterization. / Liu, Fu-Tong; Bohn, J. W.; Ferry, E. L.; Yamamoto, H.; Molinaro, C. A.; Sherman, L. A.; Klinman, N. R.; Katz, D. H.

In: Journal of Immunology, Vol. 124, No. 6, 1980, p. 2728-2737.

Research output: Contribution to journalArticle

Liu, F-T, Bohn, JW, Ferry, EL, Yamamoto, H, Molinaro, CA, Sherman, LA, Klinman, NR & Katz, DH 1980, 'Monoclonal dinitrophenyl-specific murine IgE antibody: Preparation, isolation, and characterization', Journal of Immunology, vol. 124, no. 6, pp. 2728-2737.
Liu F-T, Bohn JW, Ferry EL, Yamamoto H, Molinaro CA, Sherman LA et al. Monoclonal dinitrophenyl-specific murine IgE antibody: Preparation, isolation, and characterization. Journal of Immunology. 1980;124(6):2728-2737.
Liu, Fu-Tong ; Bohn, J. W. ; Ferry, E. L. ; Yamamoto, H. ; Molinaro, C. A. ; Sherman, L. A. ; Klinman, N. R. ; Katz, D. H. / Monoclonal dinitrophenyl-specific murine IgE antibody : Preparation, isolation, and characterization. In: Journal of Immunology. 1980 ; Vol. 124, No. 6. pp. 2728-2737.
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AU - Molinaro, C. A.

AU - Sherman, L. A.

AU - Klinman, N. R.

AU - Katz, D. H.

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