Cancerous disease modifying antibodies
一种抗体、嵌合抗体的技术，应用在结合实验领域，能够解决结合程度小等问题，达到延长存活、肿瘤负荷延长的效果
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Embodiment 1
[0064] According to the Budapest Treaty, the hybridoma cell line H460-16-2 was deposited on September 4, 2002 at the American Type Culture Collection (American Type Culture Collection), 10801 University Avenue, Manassas, Virginia, 20110-2209, and its deposit No. PTA-4621. According to 37CFR1.808, the depositor guarantees that after the grant of the patent, all restrictions on the public use of the deposited material will be permanently removed.
[0065] H460-16-2 monoclonal antibody was prepared by culturing hybridomas in CL-1000 flasks (BD Biosciences, Oakville, ON), collected and re-inoculated twice a week, and prepared according to Protein G Sepharose 4 Fast Flow (Amersham Biosciences, Antibodies were purified using standard antibody purification procedures from Baie d'Urfe, QC).
[0066] In vivo preventive tumor experiment
[0067] refer to figure 1 with 2 The data shown in , 4- to 8-week-old female SCID mice were injected subcutaneously in 100 microliters of saline so...
Embodiment 2
[0074] In vivo mature tumor assay:
[0075] 5-6 week old female SCID mice were implanted in 100 microliters of saline solution containing 5 million MB-231 breast cancer cells by subcutaneous injection into the nape of the neck. Tumor growth was measured weekly with calipers. When most of the population reaches 100mm by 34 days after transplantation 3 Tumor volume (ranging from 70-130mm 3 ), 12 mice were randomly divided into 4 treatment groups. Containing 2.7mM KCl, 1mM KH 2 PO 4 , 137mM NaCl and 20mM Na 2 HPO 4After diluting the diluent from the stock concentration, H460-16-2 or an isotype control antibody (known not to bind to MB-231 cells) was administered intravenously at a dose of 15 mg / kg / dose in a volume of 150 microliters; cisplatin was administered as A volume of 300 microliters was administered intraperitoneally at a dose of 9 mg / kg / dose (diluted with saline solution). Subsequently, a total of 10 doses of antibody were administered three times a week in the s...
Embodiment 3
[0088] Normal mouse tissue staining
[0089] The distribution of the H460-16-2 antigen was studied in mouse tissues and compared with the gp96 antigen. To determine the conditions for further experiments, IHC optimization experiments were first performed. Preparation and purification of the H460-16-2 monoclonal antibody was performed as described above.
[0090] Untreated mice implanted subcutaneously with MB-231 tumor cells were euthanized 74 days after implantation. Tissues from tumor groups and major organs were dissociated and fixed in 10% neutral buffered formalin for 48 hours. After fixation, tissues were transferred to 70% ethanol, processed, paraffin-embedded, sectioned and mounted on glass slides for staining. The slides were deparaffinized by drying in an oven at 60°C for 1 hour and deparaffinized by immersing the slides in xylene five times for 4 minutes each in a Coplin bottle. Sections were rehydrated in water after a series of graded ethanol washes (100%-75%)...
PUM
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Description
Claims
Application Information
- IPC
- A61K39/395; A61K49/00; C07K16/00; C07K16/28; C07K16/30; G01N33/569; G01N33/574
- CPC
- G01N33/57449; C07K16/3015; G01N33/57484; C07K16/00; C07K16/2884; G01N33/56972; G01N33/57415; C07K16/30
- Inventors
- 戴维·S·F·扬; 海伦·P·芬德利