ANTI-CD3e ANTIBODIES AND USES THEREOF
A technology of antibody and bispecific antibody, applied in the direction of antibody, anti-tumor drug, anti-receptor/cell surface antigen/cell surface determinant immunoglobulin, etc., which can solve the problem of limiting the available dose regimen and limiting the overall curative effect of anti-CD3 therapy And other issues
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0236] Example 1. Production of mouse anti-hCD3e antibodies
[0237] To generate anti-human CD3e (hCD3e; SEQ ID NO: 17) murine antibodies, 6-8 week old female BALB / c mice were immunized with human CD3e. through the following and figure 1 and figure 2 Anti-hCD3e antibodies were collected as indicated.
[0238] mouse immunization
[0239] 6-8 week old female BALB / c mice were immunized with His-tagged human CD3e protein at a concentration of 100 μg / ml at 20 μg / mouse. His-tagged human CD3e protein was emulsified with adjuvant and injected into four locations on the back of mice. For the first subcutaneous (s.c.) injection, the diluted antigen was emulsified with an equal volume of complete Freund's adjuvant (CFA). In subsequent subcutaneous injections, the protein was emulsified with an equal volume of incomplete Freund's adjuvant (IFA). Three days after the third injection or booster, blood (serum) was collected and analyzed for antibody titers using ELISA.
[0240] In an...
Embodiment 2
[0256] Example 2. Humanization of mouse antibodies
[0257] The starting point for humanization is mouse antibodies (eg, 10A4 and 1B1). The amino acid sequences of the heavy and light chain variable regions of these mouse antibodies were determined.
[0258] Four humanized heavy chain variable region variants (SEQ ID NOs: 21-24) and three humanized light chain variable region variants (SEQ ID NOs: 25-27) of 10A4 were constructed comprising different modification or substitution.
[0259] Four humanized heavy chain variable region variants (SEQ ID NO:28-31) and three humanized light chain variable region variants (SEQ ID NO:32-34) of 1B1 were constructed, comprising different modification or substitution.
[0260] These humanized heavy chain variable region variants can be combined with any light chain variable region variant based on the same mouse antibody. For example, 10A4-H1 (SEQ ID NO:21) can be combined with any humanized light chain variable region variant based on ...
Embodiment 3
[0262] Example 3. In vivo testing of anti-hCD3e antibodies
[0263] Experiments were performed to test the efficacy of anti-hCD3e antibodies in vivo.
[0264] A humanized CD3e mouse model was generated. A humanized CD3e mouse model was engineered to express a chimeric CD3e protein (SEQ ID NO: 20), in which a portion of the extracellular region of the mouse CD3e protein was replaced with the corresponding human CD3e extracellular region. Amino acid residues 1-126 (SEQ ID NO: 20) of the chimeric CD3e are from human CD3e (SEQ ID NO: 17). The humanized mouse model (B-hCD3e) provides a new tool for testing new therapeutic treatments in the clinical setting by significantly reducing the difference between clinical outcomes in humans and normal mice expressing mouse CD3e.
[0265] Mice were intraperitoneally injected with 1 μg or 5 μg of anti-mCD3 antibody. An equal volume of saline was injected into mice as a negative control, and tisleizumab was injected into mice for comparison...
PUM
Login to View More Abstract
Description
Claims
Application Information
Login to View More