Monoclonal antibody to human lymphotoxin and use thereof

• US 5,188,969 A
• Filed: 06/15/1989
• Issued: 02/23/1993
• Est. Priority Date: 06/20/1988
• Status: Expired due to Fees

• Current Assignee: Denki Kagaku Kogyo Kabushiki Kaisha (TDK Corporation)
• Original Assignee: Denki Kagaku Kogyo Kabushiki Kaisha (TDK Corporation)
• Inventors: Arai, Kazuhiko, Fujiwara, Akira, Motoda, Shosaku, Suzuki, Hiroyasu
• Primary Examiner(s): Kepplinger, Esther L.
• Assistant Examiner(s): Wortman, Donna C.
• Application Number:
  US07/366,568
• Time in Patent Office: 1,349 Days
• Field of Search: 436/548, 436/518, 436/824, 424/85.1, 424/88, 435/69.5, 435/70.21, 435/240.26, 435/240.27, 530/387, 530/388, 530/413, 530/809, 530/388.23, 935/81, 935/89, 935/108
• US Class Current: 436/548
• CPC Class Codes: C07K 14/525 Tumour necrosis factor [TNF]
• CPC Class Codes: C07K 16/241 Tumor Necrosis Factors
• CPC Class Codes: Y10S 436/824 Immunological separation techniques

Abstract

A monoclonal antibody reactive with a C-terminal half of human lymphotoxin, and fragments thereof; a hybridoma cell line producing a monoclonal antibody reactive with a C-terminal half of human lymphotoxin; a process for the production of a monoclonal antibody reactive with a C-terminal half of human lymphotoxin, comprising the steps of culturing the above-mentioned hybridoma in a medium to secrete the antibody, and recovering the antibody from the culture supernatant; a process for the production of a monoclonal antibody reactive with a C-terminal half of human lymphotoxin, comprising the steps of inoculating a hybridoma to a mammal, obtaining ascites from the mammal, and recovering the monoclonal antibody from the ascites; a process for the production of a hybridoma cell line producing a monoclonal antibody reactive with a C-terminal half of human lymphotoxin, comprising the step of immunizing a mammal with a conjugate of zinc and a purified recombinant human lymphotoxin, obtaining spleen cells from the immunized mammal, fusing the spleen cells with myeloma cells, and cloning a hybridoma producing the monoclonal antibody; an adsorbent for lymphotoxin comprising a solid carrier and the above-mentioned monoclonal antibody or fragment of the antibody, wherein the monoclonal antibody is bonded to the surface of the solid carrier; a process for the purification of lymphotoxin comprising the steps of placing a material containing lymphotoxin in contact with the above-mentioned adsorbent to adsorb the lymphotoxin to the adsorbent, and eluting the adsorbed lymphotoxin with an eluent; and an assay method or assay kit for lymphotoxin using the above-mentioned monoclonal antibody or fragment thereof.

Claims

1. A monoclonal antibody produced by a hybridoma selected from the group consisting of hybridoma 1-6-35 (FERM BP-1890), hybridoma 1-45-10 (FERM BP-1891) and hybridoma 1-74-54 (FERM BP-1892), or a specific binding fragment thereof.

2. A monoclonal antibody fragment according to claim 1, selected from the group consisting of Fab, and F(ab)₂.

6. An assay kit for assaying lymphotoxin, comprising a monoclonal antibody or fragment thereof according to claim 1, the monoclonal antibody or fragment thereof being attached to a solid carrier.

7. An assay kit for assaying lymphotoxin, comprising a first antibody to lymphotoxin and a second antibody to lymphotoxin, at least one of the first and second antibodies being a monoclonal antibody or a fragment thereof according to claim 1.

3. A hybridoma cell line selected from the group consisting of hybridoma 1-6-35 (FERM BP-1890), hybridoma 1-45-10 (FERM BP-1891), and hybridoma 1-74-54 (FERM BP-1892).

5. An assay method for lymphotoxin, comprising the steps of:

(a) preparing a solid carrier on which a first antibody to lymphotoxin has been attached;

(b) incubating the solid carrier from step (a) with a sample to be assayed;

(c) incubating the solid carrier from step (b) with a labeled second antibody to lymphotoxin; and

(d) detecting the label immobilized to the solid carrier,

at least one of the first and second antibodies being a monoclonal antibody or fragment thereof according to claim 3.

4. An absorbent for lymphotoxin comprising a solid carrier and a monoclonal antibody produced by a hybridoma selected from the group consisting of hybridoma 1-6-35 (FERM BP-1890), hybridoma 1-45-10 (FERM BP-1891) and hybridoma 1-74-54 (FERM BP-1892), wherein the monoclonal antibody is bonded to the surface of the solid carrier, said solid carrier selected from the group consisting of polysaccharides, synthetic polymers, and inorganic materials.