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Bispecific antibody or antibody mixture with common light chains

A technology of bispecific antibody and common light chain, which is applied in the direction of antibody, anti-receptor/cell surface antigen/cell surface determinant immunoglobulin, and the introduction of foreign genetic material using vectors, etc.

Active Publication Date: 2020-01-07
SUZHOU ALPHAMAB
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This may partly explain why Pertuzumab inhibits the growth of tumors with low HER-2 expression, while Trastuzumab binds to the extracellular IV domain of the HER-2 receptor, and dimer formation does not involve the IV domain, so Trastuzumab only effective in breast cancer patients with HER-2 overexpression

Method used

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  • Bispecific antibody or antibody mixture with common light chains
  • Bispecific antibody or antibody mixture with common light chains
  • Bispecific antibody or antibody mixture with common light chains

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Embodiment approach

[0159] CLAIMS 1. Bispecific antibody or antigen-binding part thereof, characterized in that the two light chains of said bispecific antibody or antigen-binding part thereof have the same sequence.

[0160] 2. The bispecific antibody or the antigen-binding part thereof according to embodiment 1, the heavy chain of which can correctly bind to the light chain under physiological conditions or protein expression in vitro.

[0161] 3. The bispecific antibody or antigen-binding portion thereof according to embodiment 1 or 2, wherein said light chain is capable of binding to the heavy chains of Pertuzumab and Trastuzumab, respectively.

[0162] 4. The bispecific antibody or antigen-binding portion thereof according to embodiment 3, wherein the light chain is selected from the light chain of Pertuzumab or Trastuzumab or mutants thereof.

[0163] 5. The bispecific antibody or the antigen-binding part thereof according to embodiment 4, wherein the sequence of the light chain variable re...

Embodiment 1

[0266] Embodiment 1 Acquisition of common light chain

[0267] 1. Sequence and structure acquisition

[0268] Obtain the complex crystal structure of Trastuzumab and Pertuzumab from the Protein Data Bank (PDB, www.pdb.org), the PDB number of Trastuzumab is 1N8Z, and the PDB number of Pertuzumab is 1S78 . Two screening strategies can be used to identify amino acid contacts between CH3-CH3: (i) distance of amino acid interactions (ii) solvent accessible region analysis. Here, screening is performed based on the interaction distance of amino acids.

[0269] 2. Monoclonal antibody light chain and antigen HER2 interface amino acid acquisition

[0270] According to the amino acid contact rule, interfacial amino acids refer to those amino acids whose distance between the heavy atoms of the side chain and the heavy atoms of any amino acid of another chain is less than a threshold value. Here the threshold is chosen as In some literature it is also possible to choose (Bahar and...

Embodiment 2

[0300] Example 2 Preparation and functional verification of antigenic protein HER2 variant protein

[0301] 1. Design of HER2 variant protein that only binds Pertuzumab

[0302] In 1993, Robert F. Kelley' and Mark P. O'Connell published the kinetic parameters of the binding of trastuzumab and corresponding mutations to the extracellular domain (ECD) of HER2. Among them, H91A, R50A, W95A, and Y100aA have a significant impact on the binding of trastuzumab to the extracellular domain of HER2. Hyun-Soo Cho's team crystallized the complex of trastuzumab Fab fragment and HER2 extracellular domain (ECD) in 2003 (PDB number: 1N8Z), and the results were published in Nature. By analyzing the structure of the complex of the Fab fragment of trastuzumab and the extracellular domain (ECD) of HER2 (its sequence is shown in SEQ ID NO: 18), we obtained the Fab fragment of trastuzumab and the extracellular domain of HER2 (ECD) interface contact amino acids, as shown in Table 5.

[0303] Focu...

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Abstract

The invention relates to a bispecific antibody or an antibody mixture, in particular to a bispecific antibody or an antibody mixture with common light chains, and a preparation method of the bispecific antibody or the antibody mixture. The invention further relates to a nucleic acid molecule encoding the bispecific antibody or the antibody mixture, a recombinant carrier and recombinant cell containing the nucleic acid molecule, and a detection and quantification method of the bispecific antibody or the antibody mixture. The bispecific antibody or the antibody mixture is prepared through a common light chain technology, the preparation method of the bispecific antibody or the antibody mixture is simple and controllable, mismatching of the light chains in the bispecific antibody can be avoided, as for the antibody mixture, the light chains can be expressed in the same host cell, the culture difficulty of a mixed cell group is lowered, and amplified production is better facilitated.

Description

[0001] This application is a divisional application of an invention patent application with an application date of January 8, 2015, an application number of 201510008045.8, and an invention title of "bispecific antibody or antibody mixture with a common light chain". technical field [0002] The present invention relates to a bispecific antibody or antibody mixture, and a method for preparing the bispecific antibody or antibody mixture. The present invention also relates to nucleic acid molecules encoding the bispecific antibodies or antibody mixtures, recombinant vectors and recombinant cells containing the nucleic acid molecules, and detection and quantification methods for the bispecific antibodies or antibody mixtures. Background technique [0003] Monoclonal antibody drugs have grown rapidly in the past fifteen years and have become a growth point in the pharmaceutical industry. Since 1996, a total of about 30 monoclonal antibody drugs have been approved for marketing. ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/574C12N15/12C07K14/71C07K16/46
CPCC07K14/71C12N15/63G01N33/577C07K16/32A61K2039/505A61K2039/507C07K2317/31C07K2317/515C07K2317/565C07K2317/73C07K2317/94A61P35/00C07K16/28A61K39/395C07K16/468C07K2317/52
Inventor 徐霆汪皛皛李倩逄敏洁韩莉张庆青
Owner SUZHOU ALPHAMAB
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