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Cancer-associated epitope

An epitope and adenocarcinoma technology, applied in the translation products of oncogenes, anti-receptors/cell surface antigens/cell surface determinant immunoglobulins, drug combinations, etc., can solve problems such as lack of filament elongation

Inactive Publication Date: 2005-07-13
THE SCRIPPS RES INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, lacks the extensibility of silk (13)

Method used

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preparation example Construction

[0246] The preparation of monoclonal antibodies is also routine. For example, see Kohler & Milstein, Nature, 256: 495 (1975); Coligan et al., sections 2.5.1-2.6.7; and Harlow et al., in: Antibodies: A Laboratory Manual, page 726 (Cold Spring Harbor Pub. (1988) ), the above documents are included as references in this article. A variety of well-established techniques can be used to isolate and purify monoclonal antibodies from hybridoma cultures. The separation techniques include affinity chromatography with Protein A Sepharose, size exclusion chromatography, and ion exchange chromatography. For example, see Coligan et al., sections 2.7.1-2.7.12 and sections 2.9.1-2.9.3; Barnes et al., Purification of lmmunoglobulin G (IgG), in: Methods in Molecular Biology, Vol. 10, pages 79-104 ( Humana Press (1992).

[0247] Methods of manipulating monoclonal antibodies in vitro and in vivo are well known to those skilled in the art. For example, the monoclonal antibodies used in the present inv...

Embodiment 1

[0458] Example 1: Characterization of cancer-related epitopes

[0459] Separation of COU-1 monoclonal antibody

[0460] IgM HMab, COU-1 is secreted by the hybridoma cell line B9165, which is obtained by combining the human lymphoblastoid cell line WI-L2-729-HF2 with the lymph nodes obtained from the mesenteric lymph nodes of colon cancer patients. Produced by cell fusion (35). Under aseptic conditions, mince the mesenteric lymph nodes from the tumor site of patients with colorectal cancer. The debris was removed by filtration through cotton yarn, and the lymphocytes were purified by centrifugation on Ficoll-Isopaque (Boehringer-Mannheim, Old Federal Republic of Germany and Italy).

[0461] The lymphocytes were fused with the human fusion cell line W1-L2-729-HF2 (referred to as HF2) (from Tecniclone Int., Santa Ana, Calif., USA) according to the method disclosed in the following document: Kohler, Immunological Methods Vol. II, Academic Press, 1981, pp. 285-298. Th...

Embodiment 2

[0501] Example 2: Cancer-related epitope mapping

[0502] Materials and methods

[0503] antibody

[0504] As mentioned above, IgM HMab, COU-1, is secreted by the hybridoma cell line B9165, which is obtained by combining the human lymphoblastoid cell line WI-L2-729-HF2 with the mesentery of colon cancer patients. Produced by the fusion of lymphocytes obtained in a lymph node. The hybridoma cell line B9165 was handed over to the European Cell Culture Collection (ECACC), CAMR, Salisbury, Wiltshire, SP4 OJG, UK, with the deposit number ECACC 87040201. More information about ECACC can be found on the ecacc.org website.

[0505] The human-human hybridoma cell line was grown in protein-free medium: RPMI1640 medium (Gibco, Grand Island, NY) supplemented with SSR3 serum replacement (Medicult, Copenhagen, Denmark). HMab COU-1 is purified from cell culture supernatant by affinity chromatography on agarose-conjugated murine anti-human μ-chain monoclonal antibody (Mab) (HB57, ATCC,...

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Abstract

The present invention provides a cancer-associated epitope consisting of two polypeptides, wherein the first polypeptide is derived from cytokeratin K8, and the second polypeptide is derived from cytokeratin K18. The cancer-associated epitopes become exposed during malignant transformation, especially in colon, breast, ovarian, kidney, lung and testicular tissues. Exposure of the cancer-associated epitopes occurs by cleavage and removal of the N-terminal peptides of cytokeratins K8 and K18. The invention also provides a cancer-associated epitope up to 10M-1 in cancer tissue, which is 100 times higher than that of cytokeratin K8 / K18 complex in normal tissue. The present invention provides cancer-associated epitopes, binding entities, antibodies and methods for detecting and treating cancer using the epitopes, binding entities and antibodies.

Description

Invention field [0001] The present invention relates to cancer-related epitopes, antibodies and polypeptide binding entities directed against the epitopes. The present invention also relates to a diagnostic reagent including the epitope, antibody or binding entity, and relates to the use of the epitope, antibody or binding entity for various diagnostic or therapeutic purposes. The present invention also relates to a pharmaceutical composition comprising an epitope, antibody or binding entity. Background of the invention [0002] Malignant tumors sometimes express unique antigens or "markers" that provide a means for detecting and possibly treating the cancer. For example, antigens unique to the tumor can be purified and prepared, and used to prepare antibodies. The antibody produced by the antigen can be used as a detection tool for monitoring the level of tumor markers in the host, so as to track the progress of the disease or the effect of treatment. Link a...

Claims

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Application Information

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IPC IPC(8): G01N33/483A61K38/00A61K39/00A61K39/395A61K45/00A61P35/00A61P43/00C07K14/47C07K14/82C07K16/30C12N15/09C12Q1/02G01N33/15G01N33/50G01N33/574G01N33/58
CPCA61K38/00A61K47/6843A61P35/00A61P43/00C07K14/47C07K16/18C07K2317/21C07K2317/56C07K2317/565C07K2319/00
Inventor H·迪策尔J·C·詹森纽斯
Owner THE SCRIPPS RES INST
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