Minicircle DNA carrier expressing target cell-effector cell bridge, preparation method and application thereof

A technology of effector cells and target cells, applied in the direction of intact cells/viruses/DNA/RNA components, receptors/cell surface antigens/cell surface determinants, recombinant DNA technology, etc.

Pending Publication Date: 2018-11-02
SYNO MINICIRCLE BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the existing MC.BsAb technology has deficiencies, mainly because the target range is limited and only covers the target cell surface protein antigen; and the effector cells are only limited to T cells; therefore, it is necessary to expand the scope of application of target cells and effector cells

Method used

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  • Minicircle DNA carrier expressing target cell-effector cell bridge, preparation method and application thereof
  • Minicircle DNA carrier expressing target cell-effector cell bridge, preparation method and application thereof
  • Minicircle DNA carrier expressing target cell-effector cell bridge, preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] Example 1, Construction of target cell-effector cell bridge (BTEC) bridging CD20 and CD3 (single domain antibody format)

[0056] (a) Construction of a target cell-effector cell bridge (BTEC) comprising a single-domain antibody specifically binding to CD20 (V H H), and a single domain antibody specifically binding to CD3 (V H H); CD20 is the target point on the target cells, and CD3 is the target point on the effector cells.

[0057] (b) CD20-binding single domain antibody (V H The amino acid sequence of H) is shown in SEQ ID NO: 1; the single domain antibody (V H The amino acid sequence of H) is shown in SEQ ID NO:9.

[0058] (c) Constructing a minicircle DNA vector expressing BTEC, comprising the CD20-binding single domain antibody (V H H) the coding gene, and the CD3-binding single domain antibody (V H H) coding gene; and suitable expression elements are also included in the minicircle DNA vector.

[0059] (d) An expression cassette comprising BTEC in the minic...

Embodiment 2

[0063] Example 2, Construction of target cell-effector cell bridge (BTEC) bridging CD20 and CD3 (single-chain antibody form)

[0064] (a) Construction of a target cell-effector cell bridge (BTEC), including a single-chain antibody (scFv) specifically binding to CD20, and a single-chain antibody (scFv) specifically binding to CD3; CD20 is the target on the target cell , CD3 is the target on effector cells.

[0065] (b) A CD20-binding single-chain antibody (scFv), comprising: a light chain variable region with an amino acid sequence as shown in SEQ ID NO:3, and a heavy chain variable region with an amino acid sequence as shown in SEQ ID NO:4 A CD3-binding single-chain antibody (scFv), comprising: a light chain variable region with the amino acid sequence shown in SEQ ID NO:12, and a heavy chain variable region with the amino acid sequence shown in SEQ ID NO:13.

[0066] (c) constructing a minicircle DNA carrier expressing BTEC, comprising the coding gene of the single-chain ant...

Embodiment 3

[0076] Embodiment 3, the construction of the microcircle DNA vector expressing BTEC

[0077] According to the construction ideas and methods of Examples 1 and 2, microcircle DNA vectors expressing several BTECs were constructed.

[0078] (a) Construction of BTEC, comprising single-chain T cell receptor (scTCR) binding to NY-ESO-1, and single-chain antibody (scFv) binding to CD3; the resulting minicircle DNA vector NY-ESO-1.scTCR / CD3( The structural diagram of scFv) is shown in figure 1 shown.

[0079] (b) Construction of BTEC, comprising a single-chain T cell receptor (scTCR) that binds NY-ESO-1, and a CD3-binding single-domain antibody (V H H); The resulting minicircle DNA carrier NY-ESO-1.scTCR / CD3 (V H The structure diagram of H) is as follows figure 2 shown.

[0080] (c) Construction of BTEC, comprising a single-chain T cell receptor (scTCR) that binds NY-ESO-1, and a CD16-binding single-domain antibody (V H H); The resulting minicircle DNA vector NY-ESO-1.scTCR / CD1...

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PUM

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Abstract

The invention relates to a minicircle DNA carrier expressing a target cell-effector cell bridge, a preparation method and application thereof, in particular to a minicircle DNA carrier for in vivo expression of a target cell-effector cell bridge. The target cell-effector cell bridge can specifically combine target cell effect targets and effector cell effect targets, thereby effectively connectingtarget cell to effector cell, and mediating killing to target cell by effector cell. The target cell-effector cell bridge can be used for treating cancers, infections or other diseases.

Description

technical field [0001] The present invention relates to a recombinant gene carrier in the biological field, in particular to a microcircle DNA carrier, more specifically a microcircle for expressing a target cell-effector cell bridge (the Bridge between Target and Effector Cells, BTEC for short) in vivo DNA vector. Background technique [0002] Cancer, especially malignant tumors, is an increasingly serious threat to human health. Although the body's cellular immunity plays an extremely important role in fighting against tumors, infections and other lesions, effector cells such as cytotoxic T cells and NK cells directly affect the target cells that have lesions. kill. However, in patients, immunodeficiency or immune evasion often exists, making effector cells unable to effectively recognize and kill target cells. Therefore, scientists are constantly looking for new treatments. In recent years, evidence has shown that the use of genetic engineering products target cell-eff...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/63C07K19/00C07K16/46A61P35/00A61P31/00
CPCA61K35/17C07K7/06C07K14/59C07K14/7051C07K16/2809C07K16/283C07K16/2887C07K16/468C12N15/63C07K2319/00C07K2317/73C07K2317/622C07K16/26C07K16/28C07K16/30C07K16/3069A61K2039/53C12N2800/10C07K2317/31C07K2317/569A61P31/00A61P35/00A61K48/0008A61K48/0091A61K48/0033A61K48/0075A61K48/0041A61K48/0058C07K16/2803C07K16/2869A61K39/3955C07K2317/76
Inventor 谢亦武
Owner SYNO MINICIRCLE BIOTECH CO LTD
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