Application of bone marrow mesenchymal stem cells combined with monoclonal antibodies in cancer treatment

A technology of monoclonal antibody and bone marrow mesenchyme, which is applied in the direction of antibody medical components, antibodies, anti-receptor/cell surface antigen/cell surface determinant immunoglobulin, etc., which can solve the problem of combining stem cells and therapeutic antibodies. , proliferative ability, tumor cell metastasis potential decline, etc., to achieve the effect of inhibiting proliferation and good application prospects

Inactive Publication Date: 2021-06-11
北京广未生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Decreased proliferative ability of tumor cells may lead to decreased metastatic potential of tumor cells
Although the use of bone marrow mesenchymal stem cells in the treatment of liver cancer has been studied, there are still relatively few techniques for combining stem cells with therapeutic antibodies, and further research is needed

Method used

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  • Application of bone marrow mesenchymal stem cells combined with monoclonal antibodies in cancer treatment
  • Application of bone marrow mesenchymal stem cells combined with monoclonal antibodies in cancer treatment
  • Application of bone marrow mesenchymal stem cells combined with monoclonal antibodies in cancer treatment

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Example 1 Preparation of Liver Cancer-specific Monoclonal Antibody

[0046] (1) Hepatoma Huh-7 cells and HepG2 cells cryopreserved in liquid nitrogen were thawed in a water bath at 30°C, transferred to centrifuge tubes and added with 10 ml of serum-free RPMI 1640 medium, centrifuged at 1000 r / min for 6 min, and the supernatant was discarded. The cell pellets were pipetted with RPMI1640 complete medium and then transferred to cell culture flasks respectively, and the medium was filled up to about 8 ml respectively, cultured in a 37°C, 5% CO2 incubator for 2 days, and the cells were collected for use.

[0047] The preparation method of liver cancer cell specific monoclonal antibody is as follows:

[0048] Animal immunization: select 10-week-old female Balb / c mice.

[0049] (1) Initial immunization: 150 μg of antigen (liver cancer Huh-7 cells: HepG2 cells = 1:1) and complete Freund's adjuvant at a volume ratio of 1:1 were emulsified and injected subcutaneously at multiple...

Embodiment 2

[0075] Example 2 Western blot identification of the specificity of the purified monoclonal antibody

[0076] Two kinds of liver cancer cells were lysed to extract proteins, and the lysed proteins of human epidermal cells and HeLa cells were used as controls. After reduced SDS-PAGE electrophoresis, the protein bands were transferred to PVDF membranes by 120mA constant current electrophoresis for 30 minutes. , blocked with 5% skimmed milk powder (TBST configuration, ready-to-use) at room temperature for 2 hours, discarded the blocking solution, and washed 3 times with TBST; the purified 8 kinds of monoclonal antibodies were diluted 1:250 with 5% skimmed milk powder, Incubate overnight at 4°C, wash 3 times with TBST; add HRP-labeled goat anti-mouse IgG (H+L) secondary antibody (1:5000 dilution), incubate on a shaker at room temperature for 1 hour, wash 3 times with TBST; appropriate amount of chemiluminescent solution After incubation for 5 min, the images were scanned using an A...

Embodiment 3

[0077] Example 3 Affinity determination of monoclonal antibody by SPR

[0078] Antibodies were characterized for their binding kinetics by SPR assay using a BIAcore™ T-200. Changes in surface plasmon resonance signals were analyzed to calculate association rates (kon) and dissociation rates (koff) by using a one-to-one Langmuir binding model. Equilibrium dissociation constants (KD) were calculated as the ratio koff / kon. The specific binding affinity profiles of monoclonal antibodies to liver cancer Huh-7 cells are shown in Table 2.

[0079] Table 2. Comparison of antibody binding affinities by SPR

[0080] Antibody name KD (nM) 2D3 0.23 3E2 0.56 4D3 0.29 4D6 0.31 5A3 0.58 5D7 0.45 6F4 0.62 6G5 0.34

[0081] It can be seen from Table 2 that the eight monoclonal antibodies all have good affinity.

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Abstract

The invention relates to application of bone marrow mesenchymal stem cells combined with monoclonal antibodies in cancer treatment. Monoclonal antibodies specifically aiming at the liver cancer are obtained through screening, the antibodies have a good effect of inhibiting proliferation of liver cancer cells Huh-7 and HepG2 at the same time, meanwhile it is confirmed in a mouse model test that the monoclonal antibodies have a good effect of inhibiting proliferation of tumors in a mouse body, and the antibodies and mesenchymal stem cells are combined for treating liver cancer to generate a synergistic effect, so that the antibodies have a relatively good application prospect.

Description

technical field [0001] The present invention relates to the field of biology, and more specifically, relates to the use of bone marrow mesenchymal stem cells combined with monoclonal antibodies in treating cancer. Background technique [0002] Bone marrow contains a type of non-hematopoietic stem cells, which were subsequently named mesenchymal stem cells. Initially isolated from bone marrow, it can be isolated from peripheral blood, umbilical cord blood, umbilical cord connective tissue, adipose tissue and other tissues, but bone marrow is still its main source. [0003] Bone marrow mesenchymal stem cells have self-renewal ability, have strong telomerase activity, can proliferate through asymmetric division, and maintain the number of their own stem cells. Bone marrow mesenchymal stem cells have multi-directional differentiation potential, and can differentiate into various tissue cells under certain in vivo and in vitro environments, such as osteoblasts, skeletal muscle c...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/30A61K39/395A61K35/28A61P35/00
CPCA61K35/28A61K39/39558A61P35/00C07K16/2878C07K2317/56C07K2317/73C07K2317/92A61K2300/00
Inventor 刘欢朱小明
Owner 北京广未生物科技有限公司
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