Cancerous disease modifying antibodies
A technology of antibodies and chimeric antibodies, applied in allergic diseases, antibodies, radioactive carriers, etc., can solve problems such as insufficient cancer
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Embodiment 1
[0098] Hybridoma production--hybridoma cell line AR27A807.14.1
[0099] According to the Budapest Treaty, the hybridoma cell line AR27A807.14.1 was deposited on July 18, 2006 at the National Microbiology Laboratory, Health Canada, NMLHC, the Canadian International Depository Agency, and the Bureau of Microbiology of the Canadian Ministry of Health. , Health Canada) (1015 Arlington Street, Winnipeg, Manitoba, Canada, R3E 3R2), Accession No. 180706-03. Pursuant to 37 CFR 1.808, the depositor warrants that, at the time of grant of the patent, all constraints imposed on the public availability of the deposited material are irrevocable. If the depositary cannot release a viable sample, replace the deposit.
[0100] To generate hybridomas producing anticancer antibody AR27A807.14.1, single cell suspensions of frozen human colon adenocarcinoma tumor tissue (Genomics Collaborative, Cambridge, MA) were prepared in PBS. Prepare IMMUNEASY by mixing gently TM (Qiagen, Venlo, The Nether...
Embodiment 2
[0107] in vitro binding
[0108] AR27A807.14.1 monoclonal antibody was produced by culturing hybridomas in CL-1000 flasks (BD Biosciences, Oakville, ON), harvested and reseeded twice / week. Standard antibody purification steps were followed using Protein G Sepharose 4 Fast Flow (Amersham Biosciences, Baie d'Urfe, QC). It is within the scope of the invention to use humanized, deimmunized, chimeric or murine monoclonal antibodies.
[0109] Binding of AR27A807.14.1 was assessed by cellular ELISA to: breast cancer (MDA-MB231 and MCF-7), colon cancer (HT-29, Lovo, DLD-1, SW620 and SW1116), prostate cancer (PC-3 ), pancreatic cancer (BxPC-3), and ovarian cancer (OVCAR-3), as well as non-cancer cell lines from the skin (CCD-27sk) and lung (Hs888.Lu). All cell lines were obtained from the American Type Culture Collection (ATCC; Manassas, VA). The same method as described in Example 1 was used except that purified antibody was used at a concentration of 20 μg / mL instead of hybridoma ...
Embodiment 3
[0112] In vivo tumor experiments using DLD-1 cells
[0113] Example 1 demonstrates that AR27A807.14.1 has anticancer properties against some colon cancer cell lines. refer to image 3 and 4 , 4-6 week old female SCID mice were implanted with a 5 million human colon carcinoma cell line (DLD-1 ) in 100 microliters of saline by subcutaneous injection at the nape of the neck. The mice were randomly divided into two treatment groups, 5 in each group. On the day after implantation, in a solution containing 2.7mM KCl, 1mMKH 2 PO 4 , 137mM NaCl and 20mM NaCl 2 HPO 4 After dilution of the diluent from the stock concentrate, each group was administered intraperitoneally with 20 mg / kg AR27A807.14.1 detection antibody or buffer control in a volume of 300 microliters. The antibodies and control samples were then administered in the same manner weekly for the duration of the study. Tumor growth was measured approximately every 7 days with calipers. After 6 antibody injections, the ...
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