Triabody and preparation method and applications thereof

A triple-chain antibody and antibody technology, applied in the fields of immunology and antibody engineering, can solve difficult problems

Pending Publication Date: 2019-07-05
INNOVENT BIOLOGICS (SUZHOU) CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Difficulty isolating the desired one polyspecific antibody from a complex mixture and the inherently poor yield of a theoretical maximum of 12.5% ​​make it difficult to produce four-chain Ig-like polyspecific antibodies in cell expression systems

Method used

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  • Triabody and preparation method and applications thereof
  • Triabody and preparation method and applications thereof
  • Triabody and preparation method and applications thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0264] Example 1. Construction, expression, purification, property identification and drug efficacy test of anti-CD47 / PD-L1 bispecific antibody

Embodiment 11

[0265] Example 1.1. Construction of anti-CD47 / PD-L1 bispecific antibody

[0266] In this example, two anti-CD47 / PD-L1 bispecific antibodies were constructed, named respectively (1) bispecific antibody Kh2NF-PC, and its structural schematic diagram is as follows Figure 1A shown; and (2) bispecific antibody Kh2NF-PC-NL, its structural schematic diagram is shown in Figure 1B shown.

[0267] From Figure 1A It can be seen from the schematic diagram of the structure that the bispecific antibody Kh2NF-PC is composed of 3 polypeptide chains. Peptide chain #1 has the amino acid sequence shown in SEQ ID NO: 1, which contains the sequence of SEQ ID NO: 2 derived from the anti-CD47 antibody ADI29341. VH amino acid sequence shown, CH1 amino acid sequence shown in SEQ ID NO:6 derived from human IgG1 at the C-terminal of the VH amino acid sequence, and SEQ ID NO:7 derived from human IgG1 at the C-terminal of the CH1 amino acid sequence The amino acid sequence of the Fc region is shown; t...

Embodiment 12

[0270] Example 1.2. Expression and purification of anti-CD47 / PD-L1 bispecific antibody

[0271] In this example, the nucleotide sequences encoding the three chains of the anti-CD47 / PD-L1 bispecific antibodies Kh2NF-PC and Kh2NF-PC-NL constructed in Example 1.1 were all connected to the market through multiple cloning sites The eukaryotic expression vector pTT5 sold by the company was expressed and purified in eukaryotic cells, and bispecific antibodies Kh2NF-PC and Kh2NF-PC-NL were obtained. The specific operation is as follows.

[0272] Suzhou Genewiz Biotechnology Co., Ltd. (Genewiz) was entrusted to synthesize the coding nucleotide sequences of the above three chains of the bispecific antibodies Kh2NF-PC and Kh2NF-PC-NL. The coding nucleotide sequences of the three strands synthesized were digested with restriction endonucleases XbaI (New England Biolabs) and NotI (New England Biolabs), and then combined with T4DNA ligase (New England Biolabs) respectively. The vector pTT...

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Abstract

The present invention provides a novel artificially-designed triabody comprising three polypeptide chains. A first polypeptide chain comprises a first heavy-chain variable domain, and a second polypeptide chain comprises a first light-chain variable domain, and the first heavy-chain variable domain pairs with the first light-chain variable domain to form a first antigen binding site; and a third polypeptide chain comprises a single-domain second antigen binding site and a single-domain third antigen binding site. The present invention also provides polynucleotide encoding the triabody, a vector comprising the polynucleotide, host cells comprising the polynucleotide or the vector, immunoconjugate comprising the triabody, a pharmaceutical composition comprising the triabody or the immunoconjugate, and applications of the triabody in immunotherapy, prevention and / or diagnosis of diseases.

Description

[0001] field of invention [0002] The present invention relates generally to the fields of immunology and antibody engineering. Specifically, the present invention relates to a novel artificially designed triabody, a polynucleotide encoding the triabody, a vector comprising the polynucleotide, a host cell comprising the polynucleotide or the vector , an immunoconjugate comprising the triabody, a pharmaceutical composition comprising the triabody or an immunoconjugate thereof, and the use of the triabody in the immunotherapy, prevention and / or diagnosis of diseases . [0003] Background of the invention [0004] Antibody molecules can specifically bind to their corresponding antigens, and are increasingly becoming important therapeutic, preventive and therapeutic agents for various diseases (such as cancer, autoimmune diseases, inflammatory diseases, infectious diseases, etc. and / or diagnostic agents. However, monospecific antibodies against only one target have some limitat...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/46C12N15/13C12N15/70C12N15/85C12N5/10C12N1/21A61K39/395A61P37/02A61P29/00A61P31/00A61P35/00
CPCC07K16/468C07K16/2827C07K16/2896C07K16/2803C07K16/2878C07K16/2818C07K2317/31C07K2317/92C07K2317/56A61K2039/505A61P29/00A61P31/00A61P35/00A61P37/02A61K2039/507C07K2317/52C07K2317/569C07K2317/71C07K2317/76C07K2317/94C07K2317/64C07K2319/00C07K16/46C07K2317/22C07K2317/24C07K2317/565C07K2317/626
Inventor 缪小牛匡智慧胡化静倪海晴
Owner INNOVENT BIOLOGICS (SUZHOU) CO LTD
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