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Anti-human CD73 antibody

An antibody and antibody drug technology, applied in the field of biomedicine, can solve the problems of clinical treatment limitations, no specific targeting antibody for CD73, etc.

Active Publication Date: 2021-03-26
DRAGONBOAT BIOPHARMACEUTICAL (SHANGHAI) CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no CD73-specific targeting antibody on the market
In addition, because the mouse monoclonal antibody can cause human anti-mouse antibody (HAMA) during clinical treatment, it is limited in clinical treatment

Method used

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  • Anti-human CD73 antibody
  • Anti-human CD73 antibody
  • Anti-human CD73 antibody

Examples

Experimental program
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Effect test

preparation example Construction

[0179] Antibody preparation

[0180] Any method suitable for producing monoclonal antibodies can be used to produce CD73 antibodies of the invention. For example, animals can be immunized with linked or naturally occurring CD73 protein or fragments thereof. Suitable immunization methods may be used, including adjuvants, immunostimulants, repeated booster immunizations, one or more routes may be used.

[0181] Any suitable form of CD73 can be used as an immunogen (antigen) for producing non-human antibodies specific to CD73 and screening the biological activity of the antibodies. Immunogens can be used alone or in combination with one or more immunogenicity enhancers known in the art. Immunogens can be purified from natural sources, or produced in genetically modified cells. The DNA encoding the immunogen can be genomic or non-genomic in origin (eg cDNA). DNA encoding the immunogen can be expressed using a suitable genetic vector, including but not limited to adenoviral vec...

Embodiment 1

[0223] Example 1 Preparation of anti-human CD73 monoclonal antibody

[0224] This example mainly describes the sequence of anti-human CD73 single chain antibody obtained by mouse immunization and phage display. Balb / C mice were immunized with recombinant human CD73 protein (C446, Novoprotein). After the first immunization, booster immunization was done every 14 days for a total of 4 immunizations. Mouse serum was collected for antibody titer evaluation. To evaluate the effective mice, take B cells, extract RNA for reverse transcription, and construct a phage display library. Pack the recombinant human CD73 protein at 1-5 μg / ml, add the displayed phages, and perform phage library screening. After the screening, the unwashed phages are collected and infected with host bacteria to obtain the first round of screening library. Follow this method Second and third rounds of screening. After the screening is completed, phage-ELISA identification is carried out, and those identified ...

Embodiment 2

[0233] Example 2 Affinity Detection of CQ137 Antibody

[0234] This example mainly describes the affinity detection between CQ137 and human and monkey recombinant CD73 protein, as well as the affinity detection with human CD73 positive cells (SK-OV-3, human ovarian adenocarcinoma cells) and monkey CD73 cell line (CHOK1-cynoCD73) affinity detection.

[0235] (1) Affinity with human CD73 protein

[0236] Use Biacore to measure the affinity between CQ137 and recombinant hCD73 protein, immobilize CQ137, and dilute the CD73 protein with a two-fold gradient (2.5nM-40nM). The obtained affinity diagram is shown in figure 1 .

[0237] The affinity data are shown in Table 1:

[0238] Table 1. CQ137 affinity

[0239]

[0240] The results showed that the measured affinity between CQ137 and rhCD73 was 8.653×10 -11 M.

[0241] (2) Affinity with monkey CD73 protein

[0242]Pack the recombinant monkey CD73 protein at 3 μg / ml, add CQ137 in gradient dilution, and use the ELISA method ...

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Abstract

The invention provides an anti-human CD73 antibody, and particularly provides a novel humanized antibody resistant to human CD73. The antibody can be specifically bound with a CD73 antigen and has relatively high affinity and biological activity.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular, the invention relates to an anti-CD73 humanized monoclonal antibody and its preparation. Background technique [0002] CD73, cluster of differentiation 73, also called 5'-nucleotidase, is anchored on the cell membrane through GPI (glycosylphosphatidylinositol), and its structure is a dimer. CD73 plays an important role in the tumor microenvironment. It can convert AMP into adenosine. Adenosine binds to specific G protein-coupled receptors, leading to abnormal infiltration of immune cells in tumor sites, and regulating the proliferation and migration of various cancers. It mainly inhibits the function of the immune system related to T cells, which is a sign of poor prognosis of tumors. [0003] A large number of studies in the past have shown that inhibiting CD73 can prevent tumor initiation and metastasis. Both small molecule enzyme inhibitors and macromolecular antibodies of CD73 have...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/28C07K19/00C12N15/13C12N5/10A61K39/395A61K39/00A61K47/68A61P35/00A61P35/02G01N33/68
CPCA61K39/0011A61K2039/505A61K47/6801A61P35/00A61P35/02C07K14/7051C07K16/2896C07K2317/24C07K2317/52C07K2317/56C07K2317/565C07K2317/622C07K2317/73C07K2317/92C07K2319/33C12N5/0636C12N5/0646C12N2510/00G01N33/6872G01N2333/70596
Inventor 李德彬宋作伟路娜于福涛王小燕薛玉婷林超许波琳曲平
Owner DRAGONBOAT BIOPHARMACEUTICAL (SHANGHAI) CO LTD
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