An anti-human csf-1r monoclonal antibody and its application

A technology of CSF-1R and monoclonal antibody, applied in the fields of antibody, application, anti-animal/human immunoglobulin, etc., can solve problems that need further research and development, and achieve the effect of novel sequence

Active Publication Date: 2019-05-10
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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  • An anti-human csf-1r monoclonal antibody and its application
  • An anti-human csf-1r monoclonal antibody and its application
  • An anti-human csf-1r monoclonal antibody and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Embodiment 1 obtains the mouse monoclonal antibody of specific anti-CSF1R by fusion hybridoma technology

[0036] 1.1 Animal immunity

[0037] 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 is a recombinant human CSF1R (amino acid interval Ile20-Glu 512) protein (Acrobiosystems, Cat#CSR-H5258) containing a human IgG1Fc tag at the C-terminus. Recombinant human CSF1R protein with his tag (Acro biosystems, cat#CSR-H5228) was used as the detection antigen for determination of serum titer and hybridoma screening. Briefly, remove an appropriate amount of Freund's adjuvant into a 1.5ml EP tube, and shake to mix. Prepare the antigenic protein solution with PBS. Mix the adjuvant and protein antigen solution according to the required amount, fully emulsify the antigen by pushing each other through the syringe to form a st...

Embodiment 2

[0043] Example 2 In vitro assay method for measuring CSF1R monoclonal antibody functional activity

[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) in 1xPBS to a final concentration of 2 μg / ml, add 100 μl / well to a 96-well microtiter plate, 4 degrees Pack overnight. The next day, after washing the plate 4 times with PBS solution containing 0.05% Tween 20 (ie 1xPBST), 200 μl / well of 5% skim milk powder in PBST solution was added and placed at 37°C for 2 hours to block. Wash the plate again, add 100 μl / well of diluted antibody solution or hybridoma supernatant, incubate at 37 degrees for 40 minutes, and then wash the plate 4 times. Add the prepared 60 nM 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 dilution o...

Embodiment 3

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

[0056] According to the analysis method described in Example 2, the evaluation of the binding activity of the CSF1R antibody is summarized in Table 2 below. 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 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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PUM

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Abstract

The invention relates to an anti-human CSF-1R monoclonal antibody and an application thereof. The invention also provides an encoded nucleic acid molecule of the antibody, an expression vector, a hostcell and a method of expressing the antibody. The invention also provides an antibody somatic immunoconjugate, double specific molecules, an embedded antigen acceptor, a pharmaceutical composition and an application of the antibody. Compared with the prior art, the antibody has the advantages that the human CSF-1R monoclonal antibody can be recognized specifically. The affinity of the antibody combined with CSF-1R is stronger than that of an existing anti-human CSF-1R monoclonal antibody, and the antibody is combined with a unique epitope and is novel in sequence.

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 encoding nucleic acid molecules of the antibody, expression vectors, host cells, and methods for expressing the antibody. Also provided are antibody immunoconjugates, bispecific molecules, chimeric antigen receptors, pharmaceutical compositions, and diagnostic and therapeutic methods comprising antibodies of the invention. Background technique [0002] Colony-stimulating factor 1 receptor (CSF-1R), which belongs to the type III receptor tyrosine kinase family, is encoded by the proto-oncogene c-fms. The ligand-binding domain 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 mononuclear phagocytes (Felix, et al., (2015) Structure 23, 1621-1631). And CSF1R is also highly expressed on the ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K16/28C12N15/13C12N15/63A61K39/395A61P35/00A61P29/00A61P19/02A61P9/10A61P19/08
CPCA61K2039/505A61P9/10A61P19/02A61P19/08A61P29/00A61P35/00C07K16/2866C07K2317/565C12N15/63
Inventor 陈明久谭巍
Owner BIOSION INC
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