Cancerous disease modifying antibodies
An antibody, monoclonal antibody technology, applied in the direction of antibodies, antibody medical components, anti-tumor drugs, etc., can solve problems such as weak binding
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Embodiment 1
[0111] Hybridoma preparation-hybridoma cell line AR59A816.14
[0112] According to the Budapest Treaty, the hybridoma cell line AR59A816.14 was deposited on May 17, 2005 at the International Depository of Microorganisms Canada (IDAC), located at Health Canada, 1015 Arlington Street, Winnipeg, R3E 3R2, Manitoba, Canada Ministry of Microbiology, the preservation number is 170505-02. Pursuant to 37 CFR 1.808, the depositor warrants that all restrictions on public access to the deposited material will be irrevocably removed upon grant of the patent. If the preserved specimen does not survive the deposit, the deposit should be replaced.
[0113] To prepare hybridomas producing AR59A816.14 anticancer antibody, single cell suspensions of frozen human colon tumor tissue (Genomics Collaborative, Cambridge, MA) metastasized to liver tissue were prepared in PBS. will IMMUNEASY TM (Qiagen, Venlo, Netherlands) adjuvant mixed gently and ready to use. Five to seven week old BALB / c mice w...
Embodiment 2
[0120] in vitro binding
[0121] AR59A816.14 monoclonal antibody was prepared by culturing the hybridomas in CL-1000 flasks (BD Biosciences, Oakville, ON). Supernatants were collected and re-plated twice a week. Standard antibody purification procedures were then performed using Protein G Sepharose 4 Fast Flow (Amersham Biosciences, Baie d'Urfe, QC). It is within the scope of the invention to utilize humanized, deimmunized, chimeric or murineized monoclonal antibodies.
[0122] AR59A816.14 was evaluated by flow cytometry (FACS) with breast (MDA-MB-231), colon (DLD-1, Lovo, SW620 and SW1116), prostate (PC-3) cancer cell lines, and from skin (CCD-27sk) and lung (Hs888.Lu) non-cancer cell line binding. All cell lines were obtained from the American Type Culture Collection (ATCC; Manassas, VA). Cell monolayers were first washed with DPBS (without calcium and magnesium ions) to prepare cells for flow cytometry (FACS). Cells were then detached from the cell culture plate with c...
Embodiment 3
[0125] In vivo Lovo cell tumor experiment
[0126] Examples 1 and 2 show that AR59A816.14 has anti-cancer properties on cancer cell lines and shows the highest binding activity on colon cancer cell types. The antibody was then tested in an in vivo model of human colon cancer. Referring to Figures 4 and 5, 4-6 week old female SCID mice were implanted with 1 million human colon carcinoma cells (Lovo) subcutaneously injected in 100 μl of saline at the scruff of the neck. These mice were randomly divided into two treatment groups of 5 each. On the first day after transplanting cancer cells, each group of mice was intraperitoneally administered 300 μl of AR59A816.14 test antibody (20 mg / kg) or a buffer control solution containing 2.7 mM KCl, 1 mM KH 2 PO 4 , 137mM NaCl and 20mM NaCl 2 HPO 4 The dilution solution obtained by diluting the stock solution. Then the antibody group and the control group were administered once a week in the same way for a total of 7 weeks. Tumor gr...
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