Fully humanized single-domain antibody for CD16, antigen combining fragment of antibody and applications

A single-domain antibody and binding fragment technology, applied in DNA/RNA fragments, anti-receptor/cell surface antigen/cell surface determinant immunoglobulin, application, etc., can solve the problem of short half-life

Active Publication Date: 2018-06-01
FUDAN UNIV
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Problems solved by technology

Therefore, the industry has developed an immunotherapy method that uses the activity of CD16 to treat cancer, such as bispecific molecules formed by antibodies specific to tumor surface antigens and CD16 specific antibodies on NK cells. Currently, HER2/neu and CD16 bispecific antibody, which has stronger anti-tumor activity than HER2 antibody alone; HRS-3/A9, a bispecif

Method used

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  • Fully humanized single-domain antibody for CD16, antigen combining fragment of antibody and applications
  • Fully humanized single-domain antibody for CD16, antigen combining fragment of antibody and applications

Examples

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Embodiment 1

[0093] Screening of CD16-specific single domain antibody (VH)

[0094]Using human VH 3-23 subfamily germline antibody as a template, use design and antibody engineering technology to introduce heavy chain CDR regions (CDR1, CDR2, CDR3), constructing an actual measurement library with a capacity of 150 billion (1.5×10 11 ) super-large fully human single-domain antibody library; use this phage display single-domain antibody library to screen antibodies against biotin-labeled CD16 protein; immobilize biotin-labeled CD16 protein on streptavidin-coated magnetic beads , 10 12 Antibodies displayed by each phage were incubated with 5, 4, 2, and 1 micrograms of antigen for two hours in 1, 2, 3, and 4 rounds at room temperature, and 10 phages were used for each round of screening. 12 1; use polyclonal phage ELISA to detect the enrichment of antibodies; the 1st, 2nd, 3rd, and 4th rounds of phage incubation with the coated protein, use anti-phage HRP-coupled antibodies to detect the com...

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Abstract

The invention belongs to the biotechnological field and particularly relates to a fully humanized single-domain antibody for CD16, an antigen combining fragment of the antibody and applications of theantibody and the antigen combining fragment in preparation of preparations for disease diagnosis and treatment. The invention discloses the fully humanized single-domain antibody capable of specifically combining CD16 or the antigen combining fragment of the antibody. The single-domain antibody is mainly characterized by being determined by a CDR (complementarity-determining region) specific genesequence existing in an antibody heavy chain gene variable region, and the effectively expressed antibody specifically combined with the CD16 is obtained from prokaryotic and eukaryotic cells. Different forms of genetically engineered antibodies can be modified and produced in prokaryotic and eukaryotic cells and any expression system by use of the CDR or partial or full gene of the body, and theantibody can be used for preparing preparations for clinical diagnosis or treatment of related diseases.

Description

technical field [0001] The invention belongs to the field of biotechnology, and specifically relates to fully human single-domain antibodies against CD16, antigen-binding fragments thereof, and their application in preparation of preparations for disease diagnosis and treatment. Background technique [0002] The prior art discloses that CD16 (also known as FcRγIII) is a low-affinity receptor of the Fc fragment of IgGs, which can trigger the lysis of target cells by natural killer cells (NKs) cells. Studies have shown that human NK cells contain about 15% of lymphocytes, and most NK cells have low expression of CD56 (CD56 dim ), while the expression level of CD16 was high. Human CD16 has two isoforms, CD16A and CD16B, both of which show a sequence identity of 96% in the sequence of the extracellular immunoglobulin binding region. CD16A is a transmembrane receptor expressed by macrophages, mast cells and NK cells. It co-localizes with CD3ζ and Fc-εRI-γ on NK cells. Once the ...

Claims

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Application Information

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IPC IPC(8): C07K16/28C07K19/00C12N15/13C12N15/62A61K39/395A61K45/00A61P37/04A61P29/00A61P37/08A61P35/00A61P35/04A61P35/02
CPCA61K39/39558A61K45/00A61K2039/505C07K16/283C07K2317/24C07K2317/31C07K2317/569C07K2319/30
Inventor 应天雷姜世勃吴艳玲周辰
Owner FUDAN UNIV
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