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Nucleic acid molecule and application to humanized antibody

A nucleic acid molecule and nucleotide sequence technology, applied in the field of nucleic acid molecules and their applications, can solve the problems of the small number of B-cells in the early stage, and the specificity and diversity of antibodies need to be improved, so as to reduce artificial modification and improve druggability. Effect

Pending Publication Date: 2018-09-04
CHONGQING JINMAIBO BIOTEC CO LTD
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Problems solved by technology

[0004] CN105441455A adopts the chimeric IgM sequence of human / host animals, which can realize the preparation of fully humanized antibodies, but there are still the following deficiencies: (1) the number of early B-cells of transgenic animals is less than that of normal mice; (2) The specificity and diversity of antibodies need to be improved

Method used

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  • Nucleic acid molecule and application to humanized antibody
  • Nucleic acid molecule and application to humanized antibody
  • Nucleic acid molecule and application to humanized antibody

Examples

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Effect test

Embodiment

[0061] Transfer the modified human immunoglobulin gene heavy chain vector into mice, and then immunize transgenic mice containing human immunoglobulin genes to obtain fully human antibodies. The specific steps are as follows:

[0062] 1. Construction of Immunoglobulin Gene Vectors

[0063] 1) Construction of immunoglobulin heavy chain gene (such as Figure 5 )

[0064] First, use PCR and gene synthesis to obtain the 5'-enhancer sequence of mouse IgM, all expression regulatory sequences of IgM and homology arms and screening genes (such as figure 1 ). Re-obtain human Igγ3 and Igγ1 and 3'-LCR sequences as well as homology arms and screening genes (such as image 3 ). Then all DNA fragments are homologously recombined to obtain new DNA nucleic acid molecules, and then the above-mentioned transformed DNA fragments are transferred to YAC or BAC vectors containing human immunoglobulin heavy chain genes (Ig) to construct heavy immunoglobulins. Gene clusters of chain genes such ...

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Abstract

The invention provides a nucleic acid molecule. The nucleic acid molecule comprises an immune globulin gene or a fragment of the immune globulin gene. The nucleic acid molecule is characterized by comprising an IgM gene (IgHCmu) and an IgM transduction element (switch region, Smu), and the Smu and the IgHCmu are host animal sequences. The humanoral antibody is directly acquired, the artificial modification in the later period is reduced, and the druggability of the antibody is improved. The development and the maturation of the transgenic B-cells and the number of the B-cells are completely consistent with those of the normal mouse, thereby being favorable for the differentiation of the B-cells; the specificity and the diversity of the produced antibody are improved; and the efficiency ofscreening the therapeutic antibody is improved.

Description

technical field [0001] The invention belongs to the field of biotechnology, and mainly relates to a nucleic acid molecule and its application. Background technique [0002] Fully human antibody, also known as fully human monoclonal antibody, Human (Monoclonal) Antibody, is a type of biotechnology product used in the treatment of the most important diseases such as tumors, autoimmune diseases, infectious diseases and transplant rejection. Among the global therapeutic monoclonal antibodies, 27 of the 72 monoclonal antibodies that have been marketed in Europe and the United States are human antibodies. There are two sources of human antibodies: first: the establishment of a human B cell phage antibody library, combined with phage display technology to increase the affinity; second: the use of transgenic animals to inactivate the animal's own immunoglobulin genes Incorporate part (or all) human immunoglobulin genes that have been transformed, and use the immune mechanism of ani...

Claims

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Application Information

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IPC IPC(8): C12N15/13C07K16/00C12N15/85C12N5/10
CPCC12N15/85C07K16/00C07K2317/92C07K2317/24C07K2317/21C07K2317/14C07K16/18C07K16/303C12N2015/8518A01K67/0275A01K2217/052A01K2217/15A01K2227/105A01K2267/01
Inventor 刘作华葛良鹏丁玉春邹贤刚杨松全游小燕刘雪芹吴梦
Owner CHONGQING JINMAIBO BIOTEC CO LTD
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