CS267392B1 - Mouse lymphocytar hybridom vu ifn al 6 - Google Patents

Abstract

The solution relates to murine lymphocyte antibody-producing hybridoma to human leukocyte acid-labile interferon alpha, deposited in the Virological hybridoma collection Institute of SAS in Bratislava with the designation VU IFN AL6. Monoclonal antibody the hybridoma of the VU IFN AL6 is suitable for determining the presence of acid-labile interferon alpha based on its biological inhibition activities in interferon-neutralization assays. The monoclonal antibody is also suitable for preparation mouse immunoglobulin specific sera IgG2a subclasses.

Description

(57) The present invention relates to a murine lymphocyte hybridoma producing an antibody to human leukocyte acid-labile interferon alpha, deposited in the hybridoma collection of the Institute of Virology of SAS in Bratislava under the designation VU IFN AL6. Monoclonal antibody hybridoma VU IFN AL6 is useful for determining the presence of acid-labile interferon alpha by inhibiting its biological activity in interferon-neutralization assays. The monoclonal antibody is also suitable for the preparation of mouse immunoglobulin specific sera of IgG2a subclass. EN 267392 B1 CS267 392B1 1

The present invention relates to a novel hybridoma, i.e. a hybrid unicellular organism, constructed by cell fusion of a murine myceloma lineage NS0 and a murine leucine lymphoid cell, transducing the anti-human acido antibody (a.

To date, interferon antibodies have been produced by repeatedly injecting the interferon 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 uptake, serves as a source of antibodies used primarily for quantitative or qualitative analysis of antigen - interferon molecules - in research and immunodiagnostic practice. This process, called conventional immunization, has several drawbacks. There is a heterogeneous mixture of antibodies in the serum of the immunized animals, the spectrum of which is unique in each individual organism. As a rule, in addition to antibodies to the desired antigen, the organism will produce antibodies to the antigen preparation impurities that need to be removed from the sera by saturation. Consequently, 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 ballast antibodies against impurities.

The above drawbacks of the aforementioned and hitherto used method are omitted when a hybridoma producing a monoclonal antibody against human acid-labile interferon alpha deposited in the hybridology collection of the Institute of Virology of the SAS in Bratislava, Mlýnská dolina 1, under the designation VÚ IFN AL6, is available.

Said hybridoma was obtained by a method known from the literature (Kóhler, G., 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. In: Monoclonal antibodies. Hybrids: A New Dimension in Biologica Analyzes. R. H. Kennet, T. J. McKearn, Κ. B. Bechtol, eds., New York, Plenum Press, (1980), 370) by cloning a set of hybrid cells after fusion of murine myeloma lineage NS0 and spleen cells from BALB / c mice immunized with a human leukocyte acid labile interferon alpha preparation . The advantage of a hybridoma is that it produces

Claims (1)

OBJECTIVE A mouse lyinfocyte hybridoma of the VU IFN AL6, producing a monoclonal antibody sub-gene gene, called a monoclonal antibody, is capable of specifically reacting with the appropriate interferon. The VU IFN AL6 hybridoma can be cultured in vitro in animal cell media or in vivo in the peritoneal cavity of BALB / c mice. Of the cans stored in the nitrogen liquid, the production of the antibody without further immunization with the antigens can be initiated. Example To propagate hybridoma cells in vivo, 5 10610 6 cells were injected into the peritomial 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. Ascites fluid containing the hybridoma product of VU IFN AL6 Specifically inhibited the antiviral and antiproliferative activity of 1'human acid labile interferon alpha. 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. The hybridoma cells of the VU IFN AL6 have an ultrastructural image of typical myeloma cells where the predominant organelle is free and membrane-bound polyribosomes. The basic culture medium is Dulbecco's modification of Eagle's minimal essential medium (Dulbecco, R. and Freeman, C., Virology 8/1959 /, 396). This medium, referred to as DMEM, is supplemented with gentamycin and inactivated equine serotin (10%) to cultivate the hybridoma, Elan Clona, Bratislava. The hybridoma is cultured at 37 ° C - its mean generation time is approximately 24 hours. The antibody produced is a monoclonal immunoglobulin of the IgG2a subclass. VU IFN AL6 may be industrially used as a source of anti-human leukocyte acid-labile interferon alpha antibody in quantitative or qualitative evaluation of the presence of acid-labile interferon alpha in the material under investigation or as a single subclass antibody source (IgG2a) for the preparation of the specified subclass, DISCUSSION IgG2a against human acid labile interferon alpha.