Short peptide derived from tumor antigen SAGE1

A complex, amino acid technology, used in tumor-specific antigens, tumor rejection antigen precursors, anti-tumor drugs, etc.

Active Publication Date: 2018-06-29
XLIFESC LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

After the tumor tissue is processed, the peptide fragments presented on the surface of the tumor cells can be directly identified by mass spectrometry. Although this is a complicated process, the results obtained in this way are very reliable

Method used

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  • Short peptide derived from tumor antigen SAGE1
  • Short peptide derived from tumor antigen SAGE1
  • Short peptide derived from tumor antigen SAGE1

Examples

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preparation example Construction

[0084] The preparation methods of the above TCR and antibody are known to those skilled in the art, including but not limited to, expressing and purifying from Escherichia coli cells or insect cells.

[0085] In another aspect, the present invention further provides the use of the peptides, pMHC complexes, nucleic acid molecules, vectors, cells and binding molecules of the present invention in pharmaceuticals. The peptide, pMHC complex, nucleic acid, vector, cell or binding molecule can be used to treat or prevent cancer, preferably melanoma, bladder cancer, liver cancer, epidermoid cancer, non-small cell lung cancer and squamous cell carcinoma, etc.

[0086] The present invention also provides a pharmaceutical composition, which comprises the peptide of the present invention, the pMHC complex, the nucleic acid molecule of the present invention, the cell of the present invention or the binding molecule of the present invention, and a pharmaceutically acceptable carrier. The ph...

Embodiment 1

[0093] Example 1 Identification of Polypeptides Derived from SAGE1 Antigen

[0094] Before the identification, the present invention further verified that the SAGE1 antigen is expressed in various tumor cells. Specifically, a digital single-molecule multiplex gene expression profiling system is used for detection (nanostring). The results showed that SAGE1 antigen was expressed in tumor tissues such as melanoma, bladder cancer, liver cancer, non-small cell lung cancer and squamous cell carcinoma.

[0095] The HLA-short peptide complex was purified using commercial antibody BB7.2. Specifically, the tumor cells were lysed with a buffer solution containing the non-ionic surfactant Triton X-100 (1% v / v), 1 ml of the lysate was added to 2*10^7 cells, and the tumor cells were incubated at 4° C. for 1 hour by rolling. The cell debris was removed by centrifugation, and the supernatant was first incubated with the antibody, and then rProtein A-Sepharose was added to capture the "anti...

Embodiment 2

[0102] The preparation of embodiment 2 soluble pMHC complexes

[0103] The heavy chain and light chain (β2m) of type I HLA-A*0201 molecules were expressed in E. coli (E. coli) in the form of inclusion bodies. It should be noted that the heavy chain of the HLA-A*0201 molecule used in this example does not contain its transmembrane and cytoplasmic regions in order to obtain soluble pMHC complexes. In addition, in order to facilitate the subsequent biotinylation of the soluble pMHC complex, a biotinylation tag can be added to the C-terminus of the heavy chain. The specific process of preparing the soluble pMHC complex of the present invention is as follows:

[0104] a.Purification

[0105] Collect 100ml of E.coli bacteria liquid induced to express heavy chain or light chain, centrifuge at 8000g at 4°C for 10min, wash the bacteria once with 10ml PBS, then resuspend the bacteria with 5ml BugBuster Master Mix Extraction Reagents (Merck) vigorously shake, and place in Rotate and i...

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Abstract

The invention relates to a short peptide derived from a tumor antigen SAGE1, a complex of the short peptide and an MHC molecule, and uses of the short peptide and the complex. Meanwhile, the inventionfurther relates to molecules bound with the short peptide or the complex and uses of the molecules.

Description

technical field [0001] The present invention relates to a newly discovered short peptide derived from tumor antigen SAGE1, a complex formed between the short peptide and MHC molecules and the use of the short peptide and the complex. At the same time, the present invention also relates to molecules combined with the above-mentioned short peptides or complexes, and uses of these molecules. Background technique [0002] It is well known that in many pathological conditions, such as infection, cancer, autoimmune diseases, etc., there will be inappropriate expression of some specific molecules. These molecules thus become "markers" of a pathological or abnormal state. These molecules can not only be used as markers for disease diagnosis, but also be used to produce diagnostic reagents and / or therapeutic agents. For example, markers of cancer are used to generate specific antibodies. In addition, these molecules can also effectively stimulate the specific immune response of cy...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/47C07K14/74C12N15/12C12N5/0783A61K39/00A61P35/00
CPCA61K39/00C07K14/4748C12N5/0636
Inventor 李懿余晓红龚海平刘秋萍陈琳吴万里
Owner XLIFESC LTD
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