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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Examples
Embodiment 1
[0100] Hybridoma production--hybridoma cell line AR59A367.7
[0101] According to the Budapest Treaty, the hybridoma cell line AR59A367.7 was deposited in the International Depository of Canada (the International Depository Authority ofCanada, IDAC) on February 1, 2007, and the Bureau of Microbiology, HealthCanada (Bureau of Microbiology, HealthCanada) (Canada, Ma 1015 Arlington Street, Winnipeg, Nitoba, R3E 3R2) with accession number 010207-02. 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.
[0102] To generate hybridomas producing the anticancer antibody AR59A367.7, single cell suspensions of frozen human colon metastasized liver tumor tissue (Genomics Collaborative, Cambridge, MA) were prepared in PBS. Prepare IMMUNEASY by mixing gently TM (Qiagen, Venlo, The Nether...
Embodiment 2
[0109] in vitro binding
[0110] AR59A367.7 monoclonal antibody was produced by growing 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.
[0111] The association of AR59A367.7 with breast (MDA-MB-231), colon (Lovo, DLD-1, SW620 and SW1116), prostate (PC-3), pancreas (AsPC-1 and BxPC-1) was assessed by flow cytometry (FACS). 3), a combination of lung (A549) and ovarian (OVCAR-3) carcinomas, and non-cancer cell lines from skin (CCD-27sk) and lung (Hs888.Lu). All cell lines were obtained from the American Type Tissue Collection (ATCC, Manassas, VA).
[0112] By initially using DPBS (Ca-free ++ and Mg ++ ) to wash the cell monolayer and prepare the cells for FACS. Cells were then removed fr...
Embodiment 3
[0115] In vivo tumor experiments using BxPC-3 cells
[0116] Example 1 demonstrates that AR59A367.7 has anticancer properties against human cancer cell lines. refer to Figure 4 and 5 , 8-10 week old female SCID mice were implanted with 5 million human pancreatic cancer cells (BxPC-3) in 100 microliters of PBS solution 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, 1mM KH 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 AR59A367.7 detection antibody or buffer control in a volume of 300 microliters. The antibodies and control samples were then administered weekly for the duration of the study. Tumor growth was measured approximately every 7 days with calipers. After 8 doses of antibody injections, the study was termi...
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Description
Claims
Application Information
- IPC
- C12N5/18; A61K39/395; A61K47/48; A61K51/10; A61P35/00; A61P37/04; C07K16/18; C07K16/30; C07K16/46; C07K19/00; C12P21/08; G01N33/574; G01N33/577
- CPC
- C07K16/3046; A61K51/1084; A61K51/1018; A61K2039/505; A61K51/1045; A61P35/00; A61P37/04; A61P43/00
- Inventors
- 戴维·S·F·扬; 海伦·P·芬德利