Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

CD3 resistance/CD133 resistance bi-specific antibody and CIK (cytokine-induced killer) cells loaded by CD3 resistance/CD133 resistance bi-specific antibody

A bispecific antibody and cell technology, applied in the direction of blood/immune system cells, anti-receptor/cell surface antigen/cell surface determinant immunoglobulin, anti-animal/human immunoglobulin, etc., can solve the problem of CIK cells Lack of specific targeted killing effect and limited therapeutic effect

Inactive Publication Date: 2012-11-14
黄 建华
View PDF2 Cites 10 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In adoptive immunotherapy, CIK, as a new generation of cells, has both the strong tumoricidal activity of T lymphocytes and the non-MHC-restricted tumoricidal characteristics of NK cells, and has made great progress in clinical practice, but the therapeutic effect is still limited. Is the lack of specific targeted killing of CIK cells

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • CD3 resistance/CD133 resistance bi-specific antibody and CIK (cytokine-induced killer) cells loaded by CD3 resistance/CD133 resistance bi-specific antibody
  • CD3 resistance/CD133 resistance bi-specific antibody and CIK (cytokine-induced killer) cells loaded by CD3 resistance/CD133 resistance bi-specific antibody
  • CD3 resistance/CD133 resistance bi-specific antibody and CIK (cytokine-induced killer) cells loaded by CD3 resistance/CD133 resistance bi-specific antibody

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] Example 1: Preparation of bispecific antibody

[0057] 1. Take the mouse anti-human CD3 monoclonal antibody OKT3 antibody (1 mg) and dissolve it in 500 μl of the cross-linking agent Traut's reagent (2-iminothiolane HCl; Pierce, Rockford, IL) at a 5-fold molar concentration, and act for 1 hour at room temperature; use PD- 10 column (Pharmacia, Uppsala, Sweden) was filtered to remove uncrosslinked monoclonal antibody,

[0058] 2. Take the mouse anti-human CD133 / 1 monoclonal antibody (1 mg) and dissolve it in 500 μl of the cross-linking agent Sulfo-SMCC (sulfosuccinimidyl4-(N-maleimidomethyl)cyclohexane-1-carboxylate) at a 4-fold molar concentration, and act for 1 hour at room temperature , filtered with a PD-10 column (Pharmacia, Uppsala, Sweden) to remove uncrosslinked monoclonal antibodies;

[0059] 3. After mixing the cross-linked OKT3 and the mouse anti-human CD133 / 1 monoclonal antibody overnight at 4°C, an anti-CD3 / anti-CD133 bispecific antibody was prepared.

[00...

Embodiment 2

[0062] Example 2: Equipping CIK cells with anti-CD3 / anti-CD133 bispecific antibody

[0063] 1. Culture of CIK cells

[0064] Take 10ml of peripheral venous blood from healthy volunteers, anticoagulate with heparin, dilute with normal saline 1:1, add to the upper layer of lymphocyte separation medium, collect mononuclear cells (PBMC) by centrifugation, and use RPMI1640 culture medium containing 5% fetal bovine serum Adjust the cell density to 2 x 10 5 IFN-γ was added at a final concentration of 1,000 units / mL, placed in a 5% CO2 incubator at 37°C for 24 hours, and rhIL-1α was added at a final concentration of 1000 U / ml, and the final concentration of mouse anti-human CD3 McAb was 50ng / ml, the final concentration of rhIL-2 was 1000U / ml, add rhIL-2 every 2 days, on the 14th day of culture, check the CD3, CD4, CD8 and CD56 phenotype of cultured cells by flow cytometry, and harvest CIK cells.

[0065] 2. Detection of CIK cell phenotype

[0066] Adjust the concentration of CIK ce...

Embodiment 3

[0072] Example 3: Detecting the Cytotoxicity of CIK Cells Equipped with Bispecific Antibodies to SW1990

[0073] CIK cells were used as effector cells, and the human pancreatic cancer cell line SW1990 was used as target cells to detect the killing effect of CIK cells equipped with anti-CD3 / anti-CD133 bispecific antibodies.

[0074] Take 10 samples of normal people and 10 samples of tumor patients' peripheral venous blood to prepare CIK cells, and load each case of CIK cells with anti-CD3 / anti-CD133 bispecific antibody, and compare and detect CIK cells and those loaded with anti-CD3 / anti-CD133 bispecific antibody, respectively. The killing experiment of SW1990 cells by CIK cells with specific antibodies.

[0075] The experimental procedure is as follows: take tumor cells in the logarithmic growth phase, spread 5000 cells / well on a 96-well cell culture plate, and add CIK cells and CIK cells equipped with anti-CD3 / anti-CD133 bispecific antibody prepared in Example 2 respectively ...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention relates to a bi-specific antibody, in particular to a CD3 resistance / CD133 resistance bi-specific antibody, CIK (cytokine-induced killer) cells loaded by the CD3 resistance / CD133 resistance bi-specific antibody and a preparation method of the CD3 resistance / CD133 resistance bi-specific antibody. The CD3 resistance / CD133 resistance bi-specific antibody comprises a monoclonal antibody resisting to CD3 and a monoclonal antibody resisting to CD133. The CIK cells loaded by CD3 resistance / CD133 resistance bi-specific antibody can kill and wound overexpression CD133 cancer cells in target direction and have strong effects on restraining tumor growth.

Description

technical field [0001] The present invention relates to a bispecific antibody, in particular to an anti-CD3 / anti-CD133 bispecific antibody and CIK cells loaded with the bispecific antibody. Background technique [0002] More and more evidence shows that tumor stem cells play a vital role in tumor immune escape, resistance to radiotherapy, chemotherapy, and recurrence and metastasis. Therefore, it is necessary to create new strategies to effectively target and kill tumor stem cells in order to cure tumors. Immunotherapy targeting cancer stem cells has been actively explored in the research of monoclonal antibodies and stem cell vaccines. So far, the tumor stem cell-specific markers found are very rare, and CD133 is one of the most common markers found in tumor stem cells. It exists in various tumor stem cells such as cancer, lung cancer, liver cancer and ovarian cancer, and its high expression is closely related to the low survival rate of cancer patients. [0003] Therefor...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): C07K16/46C07K16/30C07K16/28C12N5/0783
Inventor 黄建华
Owner 黄 建华
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com