CS276204B6 - Mouse hybridoma ifna1-118 producing monoclonal antibody binding the human interferon alpha 1 - Google Patents

Abstract

The purpose of the solution is to prepare a new homogeneous antibody that binds to human interferon alpha 1 and does not bind to another interferon alpha subtypes. the purpose is achieved by using a new one mouse hybridoma IFNA1-118 producing immunoglobulin monoclonal antibody G1 subclass against recombinant human interferon alpha 1. Mouse hybridoma IFNA1-118 has use in immunodiagnostics and in cleaning human interferon alpha 1.

Description

The invention relates to a novel monoclonal antibody producing mouse IFNA1-118 hybridoma that binds to human alpha 1 interferon,

Antibodies to human interferon alpha 1 are prepared in the form of antisera or monoclonal antibodies. Antisera are obtained by immunizing test animals with purified antigen (1 ' human interferon alpha 1). The antibodies thus prepared are standard and are always a mixture of antibodies with binding and neutralizing properties to human interferon alpha 1. Antisera also show cross-reactivity with related subtypes of human interferon alpha. Only hybridoma techniques for the production of monoclonal antibodies allow the production of homogeneous antibodies with exclusive binding activity to human interferon alpha 1.

Such monoclonal antibodies allow the detection of the presence of human interferon alpha 1 in a variety of biological materials by sensitive immunochemical assays. Hybridomas are obtained by a known method described in the literature / G. Kohler, C. Milstein: Nature, 256 (1975) /. The IFNA1-118 hybridoma has not been prepared yet.

The disadvantages of conventionally prepared antisera against human interferon alpha 1 are substantially eliminated by the invention, which is a murine hybridoma IFNA1-118, producing a monoclonal antibody of immunoglobulin subclass G1, which has the ability to specifically react with recombinant 1'human interferon alpha 1. Hybridoma IFNA1-118 is deposited in the Institute of Virology of SAS, Dubravska cesta 9, 842 46 Bratislava.

An advantage of the IFNA1-118 hybridoma is that it produces a homogeneous antibody that is capable of specifically reacting only with 1 ' human interferon alpha 1 and does not recognize other subtypes of human interferon alpha. After thawing, IFNA1-118 hybridoma cells continue to produce antibody without Saler immunization with human interferon alpha 1.

Example 1

Hybridomas are obtained by fusing mouse NSO myeloma cells and cells obtained from the spleen of BALB / c mice immunized with recombinant human alpha 1 interferon (ErnstBoehringer-Institut Forum Arzneimittelforschung, Vienna). Following the fusion, the IFNA1-118 hybridoma, which produces a monoclonal antibody of immunoglobulin subclass G1, selectively binding to human interferon alpha 1, was selected. The IFNA1-118 hybridoma was grown in vitro in culture media or in vivo in the peritoneal cavity of BALB / mouse mice. C. Cells are frozen and stored in liquid nitrogen and, after thawing, continue to produce the antibody without saler immunization with human interferon alpha 1. In vitro IFNA1-118 hybridoma cells as suspension culture and produce approximately 1 µg / ml specific antibody in culture medium. The culture medium is Dulbecco's modification of Eagle's Minimum Essential Medium / R. Dulbecco ,. G. Freeman: Virology B, 396 (1959)]. This medium is supplemented with inactivated horse serum (ELAN CLONE) for hybridoma culture. In order to obtain a greater amount of monoclonal antibody against human interferon alpha 1, IFNA1-118 cells were administered in an amount of 5 x 10 6 to the peritoneal cavity of BALB / c mice. The mice are pre-treated with paraffin oil (0.5 ml intraperitoneally per mouse) ten days prior to this application. 10 to 15 days after injection of IFNA1-118 cells, ascites fluid containing the desired monoclonal antibody at a concentration of approximately 1 mg / ml was collected from the mice.

The murine hybridoma IFNA1-11B is used as a source of a homogeneous antibody of immunoglobulin subclass G1, which selectively binds but does not neutralize human interferon alpha 1 and does not react with other human interferon alpha subtypes. The prepared antibody is used to detect human interferon alpha 1 in immunoassay ELISA, RIA, immunoblot and immunohistochemistry. The antibody is also suitable for immunoaffinity purification of human interferon alpha 1.

Claims (1)

Mouse IFNA1-118 hybridoma producing a monoclonal antibody of IgG1 subclass to recombinant human interferon alpha 1.