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anti-cd3 humanized antibody

A humanized antibody and antibody technology, applied in the field of biomedicine, can solve the problems of short half-life, limitations, and uncertain efficacy of bispecific antibodies

Active Publication Date: 2021-12-28
NOVOPROTEIN SCI INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Existing CD3-binding bispecific antibodies currently in clinical trials for cancer therapy are limited by short half-lives and / or variable efficacy
Although many attempts have been made to obtain anti-CD3 antibodies with particularly favorable properties, these attempts have so far had limited success

Method used

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Examples

Experimental program
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Effect test

preparation example Construction

[0312] Antibody preparation

[0313] Any method suitable for producing monoclonal antibodies can be used to produce anti-CD3 antibodies of the invention. For example, animals can be immunized with linked or naturally occurring CD3 homodimers or fragments thereof. Suitable immunization methods may be used, including adjuvants, immunostimulants, repeated booster immunizations, one or more routes may be used.

[0314] Any suitable form of CD3 can be used as an immunogen (antigen) for producing non-human antibodies specific to CD3 and screening the biological activity of the antibodies. The priming immunogen can be full-length mature human CD3, including native homodimers, or single / multiple epitope-containing peptides. Immunogens can be used alone or in combination with one or more immunogenicity enhancers known in the art. Immunogens can be purified from natural sources, or produced in genetically modified cells. The DNA encoding the immunogen can be genomic or non-genomic i...

Embodiment 1

[0354] Example 1 Production of Humanized CD3 Monoclonal Antibody

[0355] Balb / C mice were immunized with recombinant human CD3E protein (C00E, Novoprotein). After the immunization, B cells were taken to construct a phage display library, and candidate antibodies were obtained through screening. A series of humanized antibodies were obtained by structural simulation and rational design. The obtained humanized antibody heavy chain variable regions are huVH1, huVH2, huVH3 and huVH4, and the obtained humanized antibody light chain variable regions are huVL1, huVL2, huVL3 and huVL4. The VH and VL of the humanized sequence are connected by a (G4S)3 linker, and a 6HIS tag is added to the C-terminus to construct a single-chain antibody (scFv), which is expressed in E. coli and purified by a nickel column to obtain a single-chain antibody protein. The obtained humanized antibody heavy chain variable region (VH), light chain variable region (VL) sequence and CDR are as follows:

[035...

Embodiment 2

[0396] Example 2 Affinity Detection of CD3 Antibody

[0397] This example mainly describes the detection of affinity between CD3 antibody in the form of single chain antibody and human and monkey recombinant CD3E protein.

[0398] Affinity with human CD3 protein

[0399] Use ELISA to measure the affinity of CD3 antibody and recombinant hCD3 protein (CP19, Novoprotein), coat the plate with recombinant hCD3, and make 10-fold gradient dilution of CD3 antibody (starting from 20μg / ml), see figure 1 .

[0400] The calculated EC50 results are shown in Table 1.

[0401] Table 1. EC50 Affinity of CD3 Antibodies to Human CD3 Protein in ScFv Format

[0402]

[0403] 2.2 Affinity with monkey CD3 protein

[0404] The affinity between CD3 antibody and recombinant monkey CD3 protein (CW07, Novoprotein) was measured by ELISA, the recombinant monkey CD3 was coated on the plate, and the CD3 antibody was diluted 10 times (starting from 20μg / ml), see figure 2 .

[0405] The calculated E...

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Abstract

The invention provides a novel anti-CD3 humanized antibody. The anti-CD3 humanized antibody can specifically bind human and monkey CD3 proteins, and has good biological activity.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular, the invention relates to an anti-CD3 humanized antibody. Background technique [0002] Activation of T cells is critical for stimulating immune responses. T cells exhibit immune specificity and direct most cellular immune responses. Although T cells do not secrete antibodies, they are required for B lymphocytes to secrete antibodies. [0003] CD3 is a homodimeric or heterodimeric antigen expressed on T cells in association with the T cell receptor complex (TCR) and is required for T cell activation. Functional CD3 is a dimer formed by two of four different chains (ε, ζ, δ and γ). CD3 dimer arrangements include γ / ε, δ / ε, and ζ / ζ. Antibodies to CD3 have been shown to aggregate CD3 on T cells, causing T cell activation in a manner similar to the engagement of TCRs by peptide-loaded MHC molecules. Anti-CD3 antibodies therefore have therapeutic purposes involving T cell activation. In a...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K16/28C07K19/00C12N15/13C12N5/10A61K39/395A61K47/68A61P35/00A61P35/02
CPCC07K16/2809C07K14/7051A61K47/6849A61P35/00A61P35/02C07K2317/56C07K2317/565C07K2317/567C07K2317/21C07K2317/31C07K2319/00A61K2039/505
Inventor 宋作伟张清仪王英明李德彬王米秦照峰房继旋
Owner NOVOPROTEIN SCI INC
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