Humanized antibody or fragment thereof specific for cd3

A humanized antibody and specific technology, applied in specific peptides, anti-receptor/cell surface antigen/cell surface determinant immunoglobulin, allergic diseases, etc., can solve problems such as low stability

Active Publication Date: 2018-04-06
ミルテニイビオテックベーファーウントコーカーゲー
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Unfortunately, the murine OKT3 antibody shows low stability

Method used

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  • Humanized antibody or fragment thereof specific for cd3
  • Humanized antibody or fragment thereof specific for cd3
  • Humanized antibody or fragment thereof specific for cd3

Examples

Experimental program
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Embodiment approach

[0080] Antibodies or fragments thereof of the invention may be presented in various embodiments disclosed herein and described in detail in the preceding sections. All these embodiments of humanized antibodies or fragments thereof specific for the antigen CD3 have in common that in the humanized heavy and light chain variable domains respectively as SEQ ID NO: 1 to SEQ ID NO: 6 The presence of CDR1 to CDR6 is given in .

[0081] In one embodiment of the invention, the antibody or fragment thereof has a different CDR sequence (eg, IMGT, Chothia or Kabat CDRs) than the parental antibody clone OKT3. CDR sequences that differ from the murine parental antibody clone include amino acid changes, for example, substitutions of 1 or 2 amino acids in the case of CDRs of 3-4 amino acids in length, or 1, 2 in the case of CDRs of 5-7 amino acids in length. A substitution of 2, 3 or 4 amino acids, or where the length of the CDR is 10 amino acids or greater, a substitution of 1, 2, 3, 4, 5, ...

Embodiment 1

[0098] Example 1: In silico aggregation analysis of parental murine antibodies OT3 and 15E8.

[0099] The amino acid sequences of the heavy and light chain variable domains of anti-CD3 antibody clone OKT3 and anti-CD28 antibody clone 15E8 were analyzed using algorithms known in the art to predict aggregation-prone sequence regions. Such regions are known to interfere with recombinant expression of proteins, which results in low protein expression levels. Useful algorithms include, for example, PASTA (Walsh et al. (2014) Nucleic Acids Res. 42, W301-307), WALTZ (Maurer-Stroh et al. (2010) Nature Methods 7, 237-242) and AGGRESACN (Conchillo-Sole et al. (2007) ) BMC Bioinformatics 8, 65-81). Algorithms typically assign a numerical score to each amino acid, where the score is an indicator of aggregation potential. Figure 1 shows the heavy chain variable domain (SEQ ID NO: 13, Figure 1A ) and light chain variable domain (SEQ ID NO: 14, Figure 1B ) and the heavy chain variabl...

Embodiment 2

[0100] Example 2: Humanization of anti-CD28 antibody clone 15E8 by CDR grafting into human germline framework sequences.

[0101] Numbering of CDRs and amino acid residues according to IMGT nomenclature was determined by submitting the heavy and light chain sequences of the parental murine anti-CD28 antibody clone 15E8 to the IMGT "DomainGapAlign" tool. The resulting numbering plan and CDRs are defined as Figure 2A and Figure 2B shown in , and the CDRs are also described in SEQ ID NO:7 to SEQ ID NO:12. In the next step, the amino acid sequences of the light and heavy chains of the parental murine anti-CD28 antibody clone 15E8 were compared to a reference database of human germline V regions (polypeptide sequences). The current version (August 2014) of the IMGT Human IG benchmark catalog set was used as the reference database. These sequences include FR1, CDR1, FR2, CDR2 and FR3, but not CDR3 and FR4. The CDR3 of the heavy chain is mainly assembled from a small part of t...

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Abstract

The present invention provides a humanized antibody or fragment thereof specific for the antigen CD3, wherein said antibody or fragment thereof comprises a heavy chain variable domain comprising a CDR1 region of SEQ ID NO: 1, a CDR2 region of SEQ ID NO:2, and a CDR3 region of SEQ ID NO:3, and a light chain variable domain comprising a CDR1 region of SEQ ID NO:4, a CDR2 region of SEQ ID NO:5, and aCDR3 region of SEQ ID NO:6. In addition, the present invention provides a method for polyclonal stimulation of T cells comprising contacting a population of T cells with said anti-CD3 antibody, optionally with an anti-CD28 antibody, wherein said anti-CD3 antibody and optionally said anti-CD28 antibody are linked to particles. Further said anti-CD3 antibody can be used for reversible tagging of cells.

Description

technical field [0001] The present invention relates to the field of humanized antibodies or fragments thereof, in particular to antibodies or fragments thereof specific to the antigen CD3 and uses thereof. Background technique [0002] Clone OKT3 is a mouse monoclonal IgG2a specific for the human cell surface marker CD3 (T-cell surface glycoprotein CD3, a signaling co-receptor of the T-cell receptor comprising subunits γ, δ, ε, and ζ) An antibody that recognizes a small atypical region of CD3ε. OKT3 was originally described in US4658019 and Kung et al. (1979) Science 206, 347-349. For stimulation of human T cells using anti-CD3 full-length antibodies, OKT3 is well known to be a potent stimulator of T cell activation and proliferation (US6406696; US6143297; US6113901; Yannelly et al. (1990) J. Immunol. Meth 130, 91-100) . T cell activation by anti-CD3 antibodies involves complex molecular interactions. Soluble anti-CD3 antibodies do not induce T cell activation in vitro ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/28C12N5/0783C12N15/13A61K35/17
CPCA61K35/17C07K16/2809C12N5/0636C07K2317/94C07K2317/92C07K2317/24C07K16/2818C07K2317/55C07K2317/565C12N2501/51C12N2501/515A61P37/04A61P43/00C07K2317/56C07K2317/567C12N2501/599
Inventor V·诺尔H·希A·凯泽
Owner ミルテニイビオテックベーファーウントコーカーゲー
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