ovalbumin and Plasmacytoma

During the last 15 years several laboratories have attempted to generate rabbit monoclonal antibodies, mainly because rabbits recognize antigens and epitopes that are not immunogenic in mice or rats, two species from which monoclonal antibodies are usually generated. Monoclonal antibodies from rabbits could not be generated, however, because a plasmacytoma fusion partner was not available. To obtain a rabbit plasmacytoma cell line that could be used as a fusion partner we generated transgenic rabbits carrying two transgenes, c-myc and v-abl. These rabbits developed plasmacytomas, and we obtained several plasmacytoma cell lines from which we isolated hypoxanthine/aminopterin/thymidine-sensitive clones. One of these clones, when fused with spleen cells of immunized rabbits, produced stable hybridomas that secreted antibodies specific for the immunogen. The hybridomas can be cloned and propagated in nude mice, and they can be frozen without change in their ability to secrete specific monoclonal antibodies. These rabbit-rabbit hybridomas will be useful not only for production of monoclonal antibodies but also for studies of immunoglobulin gene rearrangements and isotype switching.

Topics: Animals; Antibodies, Monoclonal; Antigens; Antigens, Neoplasm; Blood Proteins; Cell Fusion; Cell Line; Enzyme-Linked Immunosorbent Assay; Epitopes; Fluorescent Antibody Technique; Humans; Hybridomas; Immunologic Techniques; Lymph Nodes; Lymphocytes; Lymphoma; Mice; Ovalbumin; Peyer's Patches; Plasmacytoma; Rabbits; Rats; Spleen; Tumor Cells, Cultured

A simplified method for producing cell hybrids by avidin-mediated electrofusion has been developed. First, biotin was attached both to immunogen and to the surface of murine myeloma cells using N-hydroxysuccinimidobiotin. Biotinylated immunogen was incubated with splenocytes derived from immunized mice and allowed to bind to surface immunoglobulins of B cells. Using streptavidin, biotinylated myeloma cells then were bridged to those B cells bearing biotinylated immunogen. Selective fusion of bridged cells was accomplished by their limited exposure to high-voltage potentials. A high frequency of hybridomas was obtained all of which secreted high titers of antibodies to a selected model immunogen, keyhole limpet hemocyanin.

Topics: Animals; Antibodies, Monoclonal; Avidin; Biotin; Cell Fusion; Electricity; Female; Hemocyanins; Hybridomas; Mice; Mice, Inbred BALB C; Ovalbumin; Plasmacytoma

Topics: Animals; Antibodies, Monoclonal; Antigen-Antibody Complex; Cell Line; Female; Intracellular Membranes; Mice; Mice, Inbred BALB C; Oocytes; Ovalbumin; Ovum; Plasmacytoma; RNA, Messenger; Xenopus

Topics: Animals; Antibodies; Antigen-Antibody Complex; Cell Line; Hybrid Cells; Mice; Mice, Inbred BALB C; Ovalbumin; Plasmacytoma; Spleen

Gamma-globulins can be crosslinked at low concentrations by simultaneous treatment with sodium sulphate and glutaraldehyde (SSG-method). This precipitate can be used for the assay of antibody or antigens. Examples are given for the measurement of (1) anti-dinitrophenyl antibody by reaction of the precipitate with 125I-dinitrophenylovalbumin, (2) IgA by inhibition of an anti-IgA precipitate with an 125I-labelled IgA myeloma protein, (3) idiotypic antibody by inhibition of the binding of a 125I-labelled myeloma protein by an anti-idiotype precipitate.

Topics: Aldehydes; Animals; Antibodies; Antigens; Binding Sites, Antibody; Cattle; Chemical Precipitation; Dinitrobenzenes; gamma-Globulins; Glutaral; Immunoassay; Immunoglobulin Allotypes; Mice; Ovalbumin; Plasmacytoma; Rabbits; Sulfates

Guinea pigs were immunized with liposomal model membranes actively sensitized with 2,4-dinitrophenyl-aminocaproylphosphatidyl-ethanolamine. The immune response was characterized by the formation of both IgM and IgG anti-DNP plaque-forming cells (PFC) which paralleled the appearance of anti-DNP antibodies in the serum. Plaque inhibition by DNP-lysine indicated that the PFC produced after liposomal immunization fell in a much narrower range of avidity groups than the PFC obtained after immunization with DNP-albumin and, in this regard, resembled MOPC 315. The restricted nature of the serum IgG anti-DNP antibodies was confirmed by isoelectric focusing and revealed the expression of a limited number of clones in all cases.

Topics: Animals; Antibody Formation; Antibody Specificity; Antibody-Producing Cells; Dinitrophenols; Erythrocytes; Female; Guinea Pigs; Hemagglutination Tests; Hemolytic Plaque Technique; Immunoglobulin G; Immunoglobulin M; Isoelectric Focusing; Liposomes; Lysine; Multiple Myeloma; Myeloma Proteins; Ovalbumin; Plasmacytoma; Serum Albumin; Sheep

Topics: Alkylation; Animals; Antibody Formation; Antigen-Antibody Complex; Cell Line; Electrophoresis, Polyacrylamide Gel; Goats; Immune Sera; Immunoglobulin A; Immunoglobulin Fragments; Leucine; Mice; Myeloma Proteins; Ovalbumin; Oxidation-Reduction; Plasmacytoma; Polymers; Sodium Dodecyl Sulfate; Tritium; Urea

Topics: Animals; Antibodies; Antigen-Antibody Complex; Antigens; Bence Jones Protein; Binding Sites; Complement Fixation Tests; Dinitrophenols; Erythrocytes; Guinea Pigs; Hemagglutination Inhibition Tests; Hemagglutination Tests; Hemocyanins; Immunoglobulin G; Mice; Neoplasm Proteins; Neoplasms, Experimental; Nitrobenzenes; Ovalbumin; Passive Cutaneous Anaphylaxis; Plasmacytoma; Protein Binding

Topics: Agammaglobulinemia; Animals; Freund's Adjuvant; gamma-Globulins; Immune Sera; Immunization; Immunoelectrophoresis; Injections, Intraperitoneal; Mice; Multiple Myeloma; Oils; Ovalbumin; Plasmacytoma; Research; Vaccination