Anti-human PD1 monoclonal antibody and application thereof

A monoclonal antibody, antibody technology, applied in the direction of antibody, anti-animal/human immunoglobulin, application, etc., can solve the problems of lack of MYPPY, inability to bind B7-1 and B7-2, etc., to achieve the effect of novel sequence

Inactive Publication Date: 2018-12-07
BIOSION INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Although PD1 is structurally similar to CTLA4, it cannot bind B7-1 and B7-2 due to the lack of MYPPY motif

Method used

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  • Anti-human PD1 monoclonal antibody and application thereof
  • Anti-human PD1 monoclonal antibody and application thereof
  • Anti-human PD1 monoclonal antibody and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Example 1 Obtaining specific anti-PD1 mouse monoclonal antibody by fusion hybridoma technology

[0040] 1.1 Animal immunity

[0041] 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 recombinant human PD1 (amino acid range Leu 25-Gln 167) protein (Acrobiosystems, Cat#PD1-H5257) containing a human IgG1 Fc tag at the C-terminus. Recombinant human PD1 protein with his s tag (Sino biological inc., cat#10377-H08H) was used as the detection antigen for serum titer determination 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 stable water-...

Embodiment 2

[0047] Example 2 In vitro analysis method for determining the functional activity of PD1 monoclonal antibody

[0048] 2.1 Determination of antibody binding ability based on capture ELSIA

[0049] 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 60nM biotin-labeled human PD1-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 dil...

Embodiment 3

[0059] Example 3 Binding activity of anti-PD1 mouse monoclonal antibody

[0060] According to the analysis method described in Example 2.1, the evaluation of the binding activity of the PD1 mouse monoclonal antibody is summarized in Table 2 below. where the reference antibody As a control, an existing commercialized anti-human PD1 monoclonal antibody was used. It can be seen from Table 2 that the binding activity of the anti-human PD1 monoclonal antibody of the present invention to human PD1 antigen is equivalent to that of the reference antibody.

[0061] Table 2 Binding activity of PD1 antibody

[0062]

[0063]

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PUM

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Abstract

The invention relates to an anti-human PD1 monoclonal antibody and application thereof, also provides coding nucleic acid molecules of the antibody, expression carriers, host cells and a method for expressing the antibody, and also provides an antibody immune coupling binder of the antibody, bispecific molecules, chimeric antigen receptors, a pharmaceutical composition and application. Compared with the prior art, the anti-human PD1 monoclonal antibody disclosed by the invention has the following advantages that the monoclonal antibody of human PD1 can be specially identified, and the bindingaffinity of the antibody and the human PD1 is stronger than that of an existing anti-human PD1 monoclonal antibody, and a sequence of the antibody is novel.

Description

technical field [0001] The present invention relates to a high-affinity and functional anti-human PD1 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] Programmed death receptor 1, also known as PD-1 or CD279, is a member of the CD28 superfamily. PD-1 is mainly expressed on activated B cells, T cells and myeloid cells (Agata et al., supra; Okazaki et al. (2002) Curr.Opin.Immunol.14:391779-82; Bennett et al. (2003 ) J Immunol 170:711-8). Unlike other members of the CD28 family, since there is no unpaired cysteine ​​in the structure of PD-1, it is suggested that PD1 mainly exists in monomeric f...

Claims

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Application Information

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IPC IPC(8): C07K16/28C12N15/13C12N15/79A61K39/395A61P35/00A61P31/00
CPCC07K16/2818A61K2039/505A61P31/00A61P35/00C07K2317/56C07K2317/565C07K2317/76C07K2317/92
Inventor 陈明久谭巍
Owner BIOSION INC
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