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Anti-cd3-binding domains and antibodies comprising them, and methods for their generation and use

An antibody and peptide-binding technology, applied in chemical instruments and methods, medical preparations containing active ingredients, antibodies, etc.

Pending Publication Date: 2020-03-24
ADIMAB LLC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

In addition, CD4+ T cell subsets include regulatory T cells, whose recruitment and expansion can potentially lead to immunosuppression and have a negative impact on long-term tumor suppression

Method used

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  • Anti-cd3-binding domains and antibodies comprising them, and methods for their generation and use
  • Anti-cd3-binding domains and antibodies comprising them, and methods for their generation and use
  • Anti-cd3-binding domains and antibodies comprising them, and methods for their generation and use

Examples

Experimental program
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Embodiment 1

[0314] Example 1: Isolation of anti-CD3 antibodies from immunized mice, and generation and isolation of humanized and optimized variants

[0315] Such as figure 1 Immunizations were performed as indicated. Briefly, in each of ten balb / c mice, 1.0 × 10 6 Individual human T cells (expressing CD3ε) and Mice were first immunized with adjuvant. Twenty-eight days after the initial immunization, in each of ten mice, 1.0 × 10 6 Individual human T cells (expressing CD3ε) and Adjuvant for the first booster. Twenty-eight days after the first immunization booster, use 1.5×10 6 T cells (expressing CD3ε) and Ten mice were given a second boost with adjuvant; however, for this second boost, five mice received human T cells (expressing CD3ε) and five mice received cynomolgus monkey (cyno) T cells ( expresses cynomolgus monkey CD3ε). Forty-two days after the second boost, all ten mice received a third boost of fifty micrograms of denatured recombinant cynomolgus CD3ε protein mixed w...

Embodiment 2

[0349] Example 2: Generation of SP34 (anti-CD3) variant antibodies: humanization and optimization of SP34

[0350] Humanization of SP34. Humanization of SP34 was performed by screening an initial panel of humanized designs produced in yeast ( Figure 27 ). Briefly, 28 designs representing combinations of four humanized VHs and seven humanized Vκ or Vλ were generated in yeast and characterized for binding to Hu CD3εδ and Cy CD3εδ Fc. Due to relatively low expression titers, five selected HuSP34 designs were generated by transient HEK and evaluated for recombinant antigen binding, cell staining and developability characteristics of these designs ( Figure 27 ). Among these designs, ADI-16606 (a combination of HC2 and LC3) was chosen with good binding and the cleanest developability profile, as shown by AC-SINS and PSR data. Notably, the background binding to CD3-expressing Jurkat cells (Jurkat CD3+), CD3-unexpressing Jurkat cells (Jurkat CD3-) and Cyno HSC-F was equivalent to...

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Abstract

Anti-CD3 binding domains and antibodies comprising them, including multispecific antibodies, with, inter alia, desirable T-cell activation and (re)directed target cell killing potency and developability, profiles are provided, as well as methods for their identification, isolation, and generation, and methods for their preparation and use. Reagents for identifying, isolating, selecting, generatingand characterizing CD3 binding domains and antibodies comprising them are also provided.

Description

[0001] Cross References to Related Applications [0002] This patent application claims priority to U.S. Provisional Application No. 62 / 503,315, filed May 8, 2017, which is incorporated herein by reference in its entirety. [0003] sequence listing [0004] This patent application contains a Sequence Listing that has been filed electronically in ASCII format, which is hereby incorporated by reference in its entirety. Said ASCII copy was created on April 7, 2017, with the name 2009186-0189_SL.TXT and a size of 2,923,505 bytes. technical field [0005] In particular, the present invention relates to anti-cluster of differentiation 3 (CD3) binding domains and antibodies comprising them (including multispecific and bispecific antibodies, and functional fragments thereof), as well as methods and reagents for their identification, isolation, preparation and use. The invention also provides reagents for identifying, isolating, selecting, producing and characterizing CD3 binding dom...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/00A61K39/395C07K16/18C07K16/28C07K16/30C07K16/32
CPCC07K16/2809A61P35/00A61P37/02A61P25/00A61P25/28C07K16/2803C07K16/30C07K16/32C07K2317/24C07K2317/31C07K2317/33C07K2317/55C07K2317/56C07K2317/565C07K2317/567C07K2317/622C07K2317/70C07K2317/73C07K2317/75C07K2317/92
Inventor L·M·沃克R·佩杰查尔E·克劳兰德M·瓦斯克斯M·W·L·梁
Owner ADIMAB LLC
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