Anti-myeloma cell polyclonal antibody and preparation method thereof
A polyclonal antibody and myeloma cell technology, applied in the direction of antibodies, anti-receptor/cell surface antigen/cell surface determinant immunoglobulin, anti-tumor drugs, etc., can solve the problem of insufficient ability to recognize myeloma cells, high myeloma Patients with limited effects and other issues, to achieve the effect of ensuring specificity and broad spectrum
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Embodiment 1
[0071] Example 1. Preparation and Identification of Rabbit Anti-Mouse or Human Whole Myeloma Cell Polyclonal Antibody
[0072] 1. Preparation of rabbit polyclonal antibody against mouse or human whole myeloma cells
[0073] Use live myeloma whole cells (mouse MPC-11 or human ARH-77) and complete Freund's adjuvant to immunize New Zealand white rabbits. After 7 to 14 days, take 1ml of blood from the ear vein of New Zealand white rabbits, and centrifuge after coagulation. Serum, the titer of polyclonal antibody detected by ELISA method was 1:2000, after that, mixed with incomplete Freund's adjuvant and living whole myeloma cells every 7 to 14 days to boost the immunization, each immunization was carried out from the edge of the ear 1ml of blood was collected from the vein, and the antibody titer was detected by ELISA method. When the antibody titer reached above 1:10000~1:50000, 80m of blood was taken from the carotid artery of each New Zealand white rabbit and filled in a glass...
Embodiment 2
[0093] Example 2 Anti-tumor effect of polyclonal antibody against myeloma cells in vivo
[0094] First, 21 Babl / c mice were inoculated intraperitoneally for 10 6 Left and right MPC-11 cells were divided into three groups after 5 days, blank control group (PBS); control antibody group (unimmunized rabbit IgG); experimental group (immunized rabbit IgG, i.e. polyclonal antibody group), 7 in each group. Then PBS, unimmunized rabbit IgG (200 μg / ml) or polyclonal antibody (200 μg / ml) were injected intraperitoneally, once every other day, a total of six times. After the sixth injection, the mice were sacrificed on the second day, and the number of intraperitoneal tumor nodules were counted by autopsy. ( Figure 5 )
[0095] see results Figure 5 , the number of abdominal nodules in mice in the control antibody and normal saline groups was significantly more than that in the antibody group of the present invention, and the number of abdominal nodules was larger, with a size range...
Embodiment 3
[0096] Example 3. In vivo anti-tumor effect of xenografted polyclonal antibodies against myeloma cells
[0097] First, 21 SCID mice were subcutaneously inoculated with 10 6 Left and right human multiple myeloma cells ARH-77 were randomly divided into three groups when tumor nodules were palpable, blank control group (NS); control antibody group (unimmunized rabbit IgG,); experimental Group (immunized rabbit IgG, polyclonal antibody group), 7 rats in each group. Then NS, 100 microliters of non-immunized rabbit IgG (200 μg / ml) or polyclonal antibody (200 μg / ml) were injected into the tail vein, once every other day, a total of six times. In order to verify the in vivo inhibitory effect of the antibody group of the present invention on xenograft tumors, the tumor volume was measured with a vernier caliper every other day, and the tumor growth curve was drawn.
[0098] see results Figure 6 , ◆, ■, ▲ respectively represent the tumor growth volume of mice treated with NS, contro...
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