Characterization of Antibodies Against Recombinant HuIFN-γ Produced by Hybridoma Cells

Balbc/c mice were immunized with purified recombinant E. coli-derived human gammainterferon (HuIFN-γ). Their spleen cells were fused with a mouse myeloma cell line (Sp2/0). Hybridomas producing antibodies reacting with HuIFN-γ were screened by a soluble-phase radioimmunoassay using pure 125I-labeled cloned IFN-γ as antigen, and tested for their ability to neutralize the antiviral activity of IFN. Three hybridomas S1-1, S1-2, and S1-3, were cloned and subcloned and remained stable. Although the antibodies produced by clones S1-1 and S1-2 were both able to neutralize specifically the antiviral activity of natural and recombinant HuIFN-γ, they appeared to recognize different epitopes on the HuIFN-γ molecule. The antibodies produced by the S1-3 clone failed to neutralize the antiviral activity of either interferon. The antibodies from all three clones were characterized as IgG1 subclass. Their affinity constants were determined from competitive inhibition curves and ranged from 1 to 4.3 × 108 M-1.

[1]  J. Kolb,et al.  SHORT COMMUNICATION: Isolation of Polyclonal Monospecific Anti HuIFN-γ Antibodies from an Antiserum Directed Against Human IFN-γ and Lymphokines , 1985 .

[2]  J. Vilček,et al.  Monoclonal antibodies to human immune interferon and their application for affinity chromatography. , 1984, Journal of immunological methods.

[3]  A. Meager,et al.  Detection of hybridomas secreting monoclonal antibodies to human gamma interferon using a rapid screening technique and specificity of certain monoclonal antibodies to gamma interferon. , 1984, Journal of interferon research.

[4]  E. Falcoff,et al.  Synergistic protection by specific antibodies and interferon against infection by Trypanosoma cruzi in vitro , 1984, European journal of immunology.

[5]  P. Kaplan,et al.  Characteristics of five monoclonal anti-alpha-fetoprotein antibodies. , 1984, Hybridoma.

[6]  E. Falcoff,et al.  Studies on purification of human gamma interferon: chromatographic behavior of accompanying IL2 and B-cell helper activity. , 1984, Journal of interferon research.

[7]  Z. Eshhar,et al.  Monoclonal antibodies to human interferon‐gamma: production, affinity purification and radioimmunoassay. , 1983, The EMBO journal.

[8]  D. Goeddel,et al.  Structure of the human immune interferon gene , 1982, Nature.

[9]  W. Degrave,et al.  Molecular cloning of human immune interferon cDNA and its expression in eukaryotic cells. , 1982, Nucleic acids research.

[10]  H. Hochkeppel,et al.  Monoclonal antibody against human IFN-γ , 1982, Nature.

[11]  M. Aguet,et al.  High affinity binding of 125I-Labeled mouse interferon to a specific cell surface receptor. II. Analysis of binding properties. , 1981, Virology.

[12]  C. Milstein,et al.  Continuous cultures of fused cells secreting antibody of predefined specificity , 1975, Nature.

[13]  K. Cantell,et al.  Quantitative studies on viral interference in suspended L cells. III. Effect of interfering viruses and interferon on the growth rate of cells. , 1962, Virology.