Cancerous disease modifying antibodies
An antibody, chimeric antibody technology, applied in allergic diseases, antibodies, radioactive carriers, etc., can solve the problem of insufficient cancer
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Embodiment 1
[0127] Hybridoma production--hybridoma cell line AR91A9.2
[0128] According to the Budapest Treaty, the hybridoma cell line AR91A9.2 was deposited in the International Depository of Canada (the International Depository Authority of Canada, IDAC) on December 5, 2006, and the Bureau of Microbiology, Health Canada (Canada , Winnipeg, Manitoba, 1015 Arlington Street, R3E 3R2), with accession number 051206-04. 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.
[0129] To generate hybridomas producing anticancer antibody AR91A9.2, single cell suspensions of frozen lung adenocarcinoma tumor tissue (Genomics Collaborative, Cambridge, MA) were prepared in PBS. Prepare IMMUNEASY by mixing gently TM(Qiagen, Venlo, The Netherlands) adjuvants were used. 5-7 week old BALB / c mice were...
Embodiment 2
[0136] in vitro binding
[0137] AR91A9.2 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 deimmunized, humanized, chimeric or murine monoclonal antibodies.
[0138] AR91A9.2 was evaluated by flow cytometry (FACS) for lung (A549, NCI-H23, NCI-H322M, NCI-H460, and NCI-H520), colon (Lovo), breast (MDA-MB-231), pancreas Combination of (BxPC-3), prostate (PC-3) and ovarian (OVCAR-3) carcinomas, and non-cancer cell lines from skin (CCD-27sk) and lung (Hs888.Lu). All cell lines, except the lung cancer cell line NCI-H322M, were obtained from the American Type Tissue Collection (ATCC, Manassas, VA). NCI-H322M was obtained from the NCI-Frederick Cancer DCTD Tumor / Cell Line Repository (Frederick, MD).
[0139] By init...
Embodiment 3
[0142] In vivo tumor experiments using NCI-H520 cells
[0143] Examples 1 and 2 demonstrate that AR91A9.2 has anticancer properties against human cancer cell lines with detectable binding to several different cancer indicators. refer to Figure 4 and 5 , 8-10 week old female SCID mice were implanted with 5 million human lung cancer cells (NCI-H520) in 100 microliters of PBS solution by subcutaneous injection at the nape of the neck. The mice were randomly divided into 2 treatment groups, 6 in each group. On the day after implantation, in a solution containing 2.7mM KCl, 1mM KH 2 PO 4 , 137mM NaCl and 20mM Na 2 HPO 4 20 mg / kg AR91A9.2 detection antibody or buffer control was administered intraperitoneally to each group in a volume of 300 microliters after dilution of the diluent from the stock concentrate. 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 day...
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