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Recombinant immunoglobulins of a new igg5 class, encoded by the human heavy chain pseudo-gamma gene

A technology of immunoglobulin and gamma gene, which is applied in the medical field and can solve problems such as the limitation of the range of changes

Pending Publication Date: 2021-01-22
UNIV DE LIMO +3
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although various mutations can be made to the structure of the IgG heavy chain to modulate its function, the range of possible changes is inherently limited by the risk of creating new immunogenic epitopes

Method used

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  • Recombinant immunoglobulins of a new igg5 class, encoded by the human heavy chain pseudo-gamma gene
  • Recombinant immunoglobulins of a new igg5 class, encoded by the human heavy chain pseudo-gamma gene
  • Recombinant immunoglobulins of a new igg5 class, encoded by the human heavy chain pseudo-gamma gene

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0180] Materials and methods

[0181] Construction of anti-CD20 IgG5

[0182] A gene synthesis reaction was used to create a fully synthetic cassette encoding anti-CD20 IgG5 (with constant sequence from the pseudo-gamma gene). The variable domains are based on the rituximab sequence. To obtain antibodies from the IgG5 class, we devised an original strategy in which:

[0183] -full length V H , DJ H Assembled exons linked to CH1, CH2 and CH3, where the exons were determined from genes not expressed in the putative human pseudo-gamma gene. This sequence is then followed by a polyadenylation site.

[0184] - Rituximab V J sequence followed by C k . This sequence is then followed by a polyadenylation site.

[0185] Finally, the construct was cloned at the HindIII and EcoRI sites into the pCDNA3.1(+) vector, which contains the neomycin resistance gene for selection.

[0186] Production

[0187] CHO and 293 cells were transfected with the plasmid vector and selected wi...

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Abstract

The present invention relates to an recombinant nucleic acid moleculecomprising an immunoglobulin heavy chain variable region (VH), an immunoglobulin light chain variable region (VL), an immunoglobulin light chain constant region (CL), an immunoglobulin heavy chain constant region (CH) corresponding to the CH1, hinge, CH2 and CHS protein domains encoded by the human pseudo-gamma gene (Psi gamma),and its uses for therapeutic or diagnostic purposes. The invention further relates to an antibody encoded by said recombinant nucleic acid molecule, and an expression cassette, vector, viral particle,host cell, transgenic organism or pharmaceutical composition comprising said recombinant nucleic acid molecule.

Description

technical field [0001] The present invention relates to the field of medicine, and in particular to novel structures of human immunoglobulins encoded by pseudogenes (ie, pseudo-gamma genes) previously thought to be unexpressed. Four immunoglobulin G classes are known, IgG1, IgG2, IgG3 and IgG4 1 (Vidarsson, G., Dekkers, G. & Rispens, T. IgG subclasses and allotypes: from structure to effector functions. Front. Immunol. 5, 520 (2014)). The present invention demonstrates an unconventional means of expressing the pseudo-gamma gene and at the same time shows that the product encoded by this pseudogene can provide a new tool for immunotherapy in humans, thereby defining an additional IgG5 class. Background technique [0002] Immunotherapy is a very active area of ​​research, with multiple developments in cancer treatment, infectious disease, and inflammatory disease. Antibodies that target tumor-specific antigens or immune checkpoints have completely changed the prognosis of ma...

Claims

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Application Information

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IPC IPC(8): C07K16/28
CPCC07K16/2887C07K2317/24C07K2317/52C07K2317/56C07K2317/622
Inventor 迈克尔·科涅
Owner UNIV DE LIMO