CS267393B1 - Mouse lymphocytar hybridoma vu ifn b13 - Google Patents

Abstract

The solution relates to murine lymphocyte 1-human antibody-producing hybridoma fibroblast interferon beta, deposited in the Virological Hybridoma Collection Institute of SAS in Bratislava under the designation of IFI B13. Hybridoma monoclonal antibody The IFN B13 is suitable for staging inhibition of interferon beta its biological activity in interferon-neutralizing tests. Monoclonal antibody it is also suitable for the preparation of mouse-specific sera immunoglobulin IgG1 subclass.

Description

(57) The present invention relates to a murine lymphocyte hybridoma producing an antibody to human fibroblast interferon beta deposited in the hybridoma collection of the Institute of Virology of SAS in Bratislava under the designation of IFI B13. Monoclonal antibody hybridoma VU IFN B13 is useful for determining the presence of interferon beta based on inhibition of its biological activity in interferon-neutralization assays. The monoclonal antibody is also suitable for the preparation of sera specific for mouse immunoglobulin IgG1 subclass. EN 267393 B1 EN 267 393 B1 1

The present invention relates to a novel hybridoma, i.e., a hybrid unicellular organism, or a cell fusion of a murine myeloma lineage NS0 and a mouse spleen lymphoid cell, which crosses the antibody in human inlerferon beta.

To date, interferon beta antibodies are produced by repeating interferon repeated injection as an antigen to experimental animals, most often rabbits or sheep. The serum of the animals so immunized, taken after a certain period of antigen exposure, serves as a source of antibodies used primarily for quantitative or qualitative analysis of the interferon beta antigen in research and immunodiagnostic practice. This process, called conventional immunization, has several drawbacks. There is a heterogeneous mixture of antibodies in the serum of immunized animals, the spectrum of which is unique and unrepeatable in each individual organism. As a rule, in addition to antibodies to the desired antigen, the organism creates antibodies to the antigen preparation impurities that are needed to remove from the sera by saturation. Consequently, production batches of conventional sera are difficult to standardize and are in a wide range of quality. To produce each batch, a clean immunization antigen and other antigens need to be prepared to saturate the anti-dirt ballast antibodies.

The aforementioned drawbacks of the aforementioned and hitherto used method are omitted when a hybridoma producing a monoclonal human interferon beta antibody deposited in the Hybridoma Collection of the Institute of Virology of the SAS in Bratislava, Mlýnská dolina 1, under the designation VÚ IFN B13, is available.

Said hybridoma was obtained by a method known from the literature (Kóhler, C., Milstein, C .: Continuous cultures of fused cells secreting antibody predefined specificity. Nature, 256, / 1975 /, 495; Gerhard, W .: Fusion of cells in suspension and outgrowth of hybrids in conditioned medium: Hydrids: A New Dimension in Biological Analyzes, RH Kennt, TJ McKearn, B. Bechtol, eds., New York, Plenum Press, / 1980/370 by cloning a set of hybrid cells after fusion of mouse myeloma line NSO and spleen cells of BALB / c mice immunized with human fibroblast interferon beta preparation. The advantage of a hybridoma is that it produces

Claims (1)

OBJECTIVE A mouse lymphocyte hybridoma VIFFNB13 producing a monoclonal antibody subclass 2 antibody gene, the so-called monoclonal antibody, is capable of specifically reacting with interferon beta. The V1FNB13 hybridoma can be cultured in vitro in media suitable for animal cells, or in vivo in the peritoneal cavity of BALB / c mice. Of the canned foods stored in liquid nitrogen, production of the antibody may be initiated without further antigen immunization. Example To propagate hybridoma cells in vivo, 5 χ 10 ® cells were injected into the peritoneal cavity of mice. For better attachment of the applied cells, the mouse was pre-treated with paraffin oil (0.5 ml intraperitoneally) 15 days before hybridoma cell transfer. After 20 days of hybridoma growth in the peritoneal cavity, the mouse was killed and the produced ascites fluid collected. A total of 7 ml of ascitic fluid was obtained which contained 8 mg / ml immunoglobulin. Ascitic fluid containing the hybridoma product of VU IFN B13 specifically inhibited the antiviral and antiproliferative activity of human human interferon beta. In vitro growths of hybridomas as a semi-suspension culture. They are shaped (round) and the size characteristic of myeloma cells, containing fused cell nuclei, are aneuploid. IFN B13 hybridoma cells have an ultrastructural image of typical myeloma cells, where free and membrane-bound polyribosomes are the predominant organelle. The basic culture medium is Dulbecco's modification of Eagle's minimal essential medium (Dulbecco, R. and Freeman, G., Virology 8/1959 /, 396). This medium, referred to as DMEM, is supplemented with gentamycin and inactivated horse serum (10%, Elan clone, Bratislava) to cultivate the hybridoma. The hybridoma is cultured at 37 ° C - its mean generation time is about 24 hours. The antibody produced is a monoclonal immunoglobulin of the IgG1 subclass. The IFU B13 VU hybridoma can be industrially used as a source of anti-human fibroblast interferon beta antibody in quantitative or qualitative evaluation of the presence of interferon beta in the investigated material or as a single subclass (IgG1) antibody source for the subclass specific sera. DISCUSSION of IgG1 against human interferon beta.