CS269049B1 - Mouse lymphocyte hybrid VU 315/4 - Google Patents

Abstract

The solution concerns the present lymphocyte hybridoma, producing an IgG 1 subclass antibody against the nucleoprotein of the influenza A virus. The hybridoma is stored in the hybridoma collection of the Institute of Virology of the Slovak Academy of Sciences in Břetislava under the designation VU 315/4.

Description

CS 269 049 B1 1

The invention aa relates to a mouse lymphocyte hybridoma, i.e. a hybrid single-cell organization, constructed by cellular fusion of a mouse Sp2 / 0a myeloma line, and a mouse spleen cell producing an influenza A virus antibody,

Antibodies to influenza virus proteins have so far been prepared by virus or virus. its isolated proteins are repeatedly injected as the antigen of the test animals most frequently in rabbits, mice, or ferrets. The serum of such immunized animals, taken after a certain time of antigen uptake, serves as a source of antibodies used primarily for the quantitative or qualitative analysis of the antigen - influenza virus - in research and immunodiagnostic practice.

This process, termed conventional immunization, has several disadvantages. In sero-immunized animals, there is a heterogeneous mixture of antibodies, the spectrum of which is spicy and unrepeatable in each individual organism. In addition to antibodies to the antigen of interest, the organism forms an antibody to an impurity mantan preparation that is desaturated from the sera, so that batches of conventional sera can be standardized and have a wide variety of properties. For the production of each batch, a clean immunization antigen needs to be prepared by additional antigens to saturate the ballast counterparts against dirt.

The aforementioned drawbacks of the aforementioned and hitherto used process waste, if available, are the hybrids producing the monoclonal antibody against the influenza A virus nucleoprotein deposited in the hybrid collection of the Virological Institute of SAS in Bratislava, Mlynska dolina 1. under the designation VU 315/4,

Said hybridoma was obtained by a method known from the literature (KdnlerG, Milstein. C, Continuous cultures of fused cells secreting antibodies of pre-defined specificity. Nature, 256, (1975), 495: Gerhard W .: Fusion of cells insuspension) and autgrowth of hybrids in conditloned medium, In; Monoclonal anti-dies, Hybrids; and New Oimension in Biological Analyzes, RH Kennet, T.3 McKearn, KB Bechtol, eds., New York, Plenum Press (1980), 370 .) cloning of the hybrid cell set after fusion of mouse myeloma line 3p2 / 0 and cells obtained from the spleen of BALB / c mice immunized with influenza A / Dunedin / 4/73 (H3N2) virus. An advantage of the hybrid of the invention is that it produces a hemeogenic antibody, a so-called monoclonal antibody that is capable of specifically reacting with the influenza A virus nucleoprotein, the VU 315/4 hybrid can be cultured in animal cells suitable for animal cells, or in vivo in the peritoneal BALB / c mouse cavity, 2 cans, stored in liquid nitrogen, production of antibody without immunization with the antigen can be initiated. Example

In order to propagate hybridoma cells in vivo, 5 x 10 6 cells were injected into the peritoneal cavity of the mouse. For better attachment of the applied cells to bolamus 15 days prior to cell hybridoma transfer, paraffin oil (0.5 ml intraperitoneal), after 20 days of growth by hybridoma in the peritoneal cavity bolams killed and produced ascites fluid collected, a total of 8 mlascitic fluid containing 5 mg / ml was obtained immunoglobulin. Ascitic fluid containing laundry hybrid VU 315/4 showed specific binding to influenza A viruses (e.g., H1, H2, H3 subtype) in radioimmunoassay and not binding to B, and bound to nucleoprotein in radioimmunoassay influenza virus A.

Claims (1)

SUBJECT OF THE INVENTION Mouse lymphocyte hybrids VU 315/4, producing a monoclonal antibody of the IgG 1 subclass against the nucleoprotein of the influenza A virus.