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Chimeric antibody and immunocyte

A technology of chimeric antibody and antibody, which is applied in the direction of anti-animal/human immunoglobulin, anti-receptor/cell surface antigen/cell surface determinant immunoglobulin, cells modified by introducing foreign genetic material, etc. No single-domain antibody targeted therapy and other problems have been seen, and the effect of strong specificity and high killing rate has been achieved

Inactive Publication Date: 2013-05-08
INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Currently, there is only one document reporting the use of immunotoxins based on Fab antibodies specific to HLA-A2 / gp100-peptide complexes for the immunotherapy of melanoma. There are no reports of targeted therapy based on domain antibodies, and no reports on cell-targeted therapy based on antibodies specific to HLA-A2 / gp100-peptide complexes

Method used

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  • Chimeric antibody and immunocyte
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  • Chimeric antibody and immunocyte

Examples

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Embodiment 1

[0051] Example 1, HLA-A 2 / gp100 209-217(215E)(IMDQVPESV) Preparation and functional verification of specific single domain antibody (GPAT)

[0052] NK-92-MI cells were purchased from ATCC with catalog number CRL-2408; T2 cells were purchased from ATCC with catalog number CRL-1992 TM ; Human melanoma cells Malme-3m were purchased from ATCC, catalog number HTB64.

[0053] Human NK-92-MI cell line and human T lymphoma / B cell hybrid cell T2 (HLA-A2 + and gp100 - ) were cultured with 1640 complete medium containing 10% fetal bovine serum, 100 U / mL penicillin and 100 μg / mL streptomycin.

[0054] NK-92-MI-GPAT-ζ cells were cultured with 1640 complete medium containing 1000 μg / ml G418.

[0055] Human melanoma cell line Mel-624 (HLA-A2 + and gp100 + ) and Malme-3m (HLA-A2 + and gp100 + ) were cultured with DMEM complete medium containing 10% fetal bovine serum, 100 U / mL penicillin and 100 μg / mL streptomycin.

[0056] 1. Preparation of coding gene of single domain antibody (G...

Embodiment 2

[0089] Example 2, Construction and Application of Chimeric Antibody GPAT-ζ

[0090] One, the construction of the recombinant vector containing the coding gene of chimeric antibody GPAT-ζ

[0091] The gene encoding the GPAT-ζ chimeric antibody is composed of HLA-A2 / gp100 209-217(215E)(IMDQVPESV) The coding gene of the specific single domain antibody GPAT ( Figure 5 GPAT in A), the sequence encoding Myc tag (EQKLISEEDL) ( Figure 5 Myc tag in A), human HLA-A 2 The gene encoding the transmembrane region fragment (amino acid 291-340) of the molecule ( Figure 5 TM in A, i.e. the following a2tm), the coding gene of the intracellular region fragment (amino acid 52-164) of human CD3 complex protein ζ chain ( Figure 5 ζ) in A is fused, and its structure is as follows Figure 5 As shown in A.

[0092] hHLA-A 2 The amino acid sequence of the transmembrane region fragment of the molecule can be specifically shown as sequence 3 in the sequence listing; human HLA-A 2 The nucleoti...

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Abstract

The invention discloses a chimeric antibody and an immunocyte. The chimeric antibody provided by the invention has an amino acid sequence containing a target antibody, a transmembrane domain segment of a HLA-A2 (Human Leukocyte Antigen-A2) molecule and an intracellular domain segment of a CD3 compound protein zeta chain in sequence from a terminal N to a terminal C. The chimeric antibody can be expressed on the surfaces of any cells, when the chimeric antibody is expressed on the surface of an immunocyte, the immunocyte is endowed with a specific immunity by virtue of the target antibody on the chimeric antibody, and thus, the chimeric antibody provided by the invention can be specifically combined with a plurality of antigens along with different target antibodies in the chimeric antibody. The chimeric antibody, a single-domain antibody and an immunocyte for expressing the chimeric antibody, provided by the invention, are about to have broad application prospects in the cellular immunotherapy.

Description

technical field [0001] The invention relates to a chimeric antibody and immune cells. Background technique [0002] Natural killer cells (Natural killer, NK) are a subtype of lymphocytes, which play an important role in the host's cellular immune response to malignant tumors. The killing of NK cells on tumors is not MHC-restricted, non-specific, and does not require antigen pre-sensitization. Many cytokines (including IL-2) can enhance the tumoricidal activity of NK cells. The function of NK cells is regulated by activating and inhibitory receptors expressed on their cell surface. [0003] Studies have shown that NK cells can effectively kill tumor cells with down-regulated MHC-I molecules, which makes them effective in killing tumor mutant cells that escape immune response. Many studies have shown that activated autologous or allogeneic NK cells cultured in vitro can effectively inhibit the growth of tumors in mice by releasing perforin and cytokines. However, the isola...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K19/00C07K16/28C12N15/13C12N15/62C12N15/63C12N1/00C12N5/10
Inventor 高斌刘容枝朱学楷
Owner INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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