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A kind of anti-human csf-1r monoclonal antibody and its application

A CSF-1R and monoclonal antibody technology, applied in the direction of antibodies, anti-animal/human immunoglobulins, applications, etc., can solve problems that need further research and development, and achieve the effect of novel sequences

Active Publication Date: 2019-07-23
BIOSION INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Although CSF1R antibodies have been developed, CSF1R inhibitors with higher affinity and other key properties of drugs for the treatment of the above diseases, especially monoclonal antibodies, still need to be further researched and developed

Method used

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  • A kind of anti-human csf-1r monoclonal antibody and its application
  • A kind of anti-human csf-1r monoclonal antibody and its application
  • A kind of anti-human csf-1r monoclonal antibody and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Example 1 Obtaining specific anti-CSF1R mouse monoclonal antibodies by fusion hybridoma technology

[0036] 1.1 Animal immunity

[0037] The mice were immunized according to the general method in the literature (E Harlow, D. Lane, Antibody: A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., 1998). The immunogen was a recombinant human CSF1R (amino acid interval Ile20-Glu 512) protein (Acrobiosystems, Cat#CSR-H5258) containing a human IgG1Fc tag at the C-terminus. The recombinant human CSF1R protein with his s tag (Acro biosystems, cat#CSR-H5228) was used as the detection antigen for serum titer determination and hybridoma screening. Briefly, take out an appropriate amount of Freund's adjuvant into a 1.5ml EP tube and mix it with a shaker. Prepare the antigen protein solution with PBS. Mix the adjuvant and protein antigen solution according to the required amount, push each other through a syringe to fully emulsify the antigen to form a sta...

Embodiment 2

[0043] Example 2 In vitro analytical method for determining the functional activity of CSF1R monoclonal antibody

[0044] 2.1 Determination of antibody binding ability based on capture ELSIA

[0045] Prepare goat anti-mouse IgG Fcr-specific secondary antibody (Jackson Immuno Research, #115-006-071) with 1xPBS to a final concentration of 2μg / ml, add 100μl / well to 96-well microtiter plate, 4 degrees Coated overnight. The next day, after washing the plate 4 times with a PBS solution containing 0.05% Tween 20 (ie 1xPBST), 200 μl / well of 5% skimmed milk powder in PBST solution was added and placed at 37°C for 2 hours. Wash the plate again, add 100μl / well of diluted antibody solution or hybridoma supernatant, incubate at 37°C for 40 minutes, and then wash the plate 4 times. Add the prepared 60nM biotin-labeled human CSF1R Fc protein solution (in 2.5% skimmed milk powder in PBST) at 100 μl / well, incubate at 37°C for 40 minutes, and then wash the plate 4 times. Add 1:10000 diluted horse...

Embodiment 3

[0055] Example 3 Binding activity of anti-CSF1R mouse monoclonal antibody

[0056] According to the analysis method described in Example 2, the binding activity of the CSF1R antibody tested is summarized in Table 2. Among them, Benchmark (Cabiralizumab) is used as a control, which is an existing non-commercialized anti-human CSF1R monoclonal antibody. It can be seen from Table 2 that the binding activity of the anti-human CSF-1R monoclonal antibody of the present invention to the human CSF1R antigen is significantly better than that of Benchmark.

[0057] Table 2 Binding activity of purified CSF1R mouse monoclonal antibody

[0058]

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Abstract

The invention relates to an anti-human CSF-1R monoclonal antibody and application. Meanwhile, the invention provides a coding nucleic acid molecule of the antibody, an expression vector, a host cell and a method which is used for expressing the antibody. The invention further provides an antibody immunoconjugate, a bi-specific molecule, a chimeric antigen receptor, a medicine compound and application of the antibody. Compared with the prior art, the anti-human CSF-1R monoclonal antibody has the following advantages that the monoclonal antibody can specifically recognize human CSF-1R, the binding affinity of the antibody and the human CSF-1R is higher than those of existing anti-human CSF-1R monoclonal antibodies, sequences of the anti-human CSF-1R monoclonal antibody are novel, and the anti-human CSF-1R monoclonal antibody bonds a unique epitope.

Description

Technical field [0001] The invention relates to a high-affinity and functional anti-human CSF1R monoclonal antibody or antibody fragment. The present invention also provides the antibody-encoding nucleic acid molecule, expression vector, host cell, and method for expressing the antibody. Also provided are antibody immunoconjugate conjugates, bispecific molecules, chimeric antigen receptors, pharmaceutical compositions, and diagnostic and therapeutic methods including the antibodies of the present invention. Background technique [0002] Colony stimulating factor 1 receptor (CSF-1R), belonging to the type III receptor tyrosine kinase family, is encoded by the proto-oncogene c-fms. The protein structure contains the intracellular kinase domain and the extracellular 5 immunoglobulins. A ligand binding region composed of a protein-like structure. CS1R is expressed on the surface of mononuclear phagocytes and plays a key role in the survival, proliferation and differentiation of mon...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K16/28C12N15/13A61K39/395A61P35/00A61P29/00A61P19/02A61P9/10A61P19/00
CPCA61P9/10A61P19/00A61P19/02A61P29/00A61P35/00C07K16/2866C07K2317/56C07K2317/565C07K2317/76C07K2317/92
Inventor 陈明久谭巍
Owner BIOSION INC
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