Tumour antigen protein and tumour antigen peptide

A tumor antigen and protein technology, applied in the direction of peptide/protein components, antineoplastic drugs, peptide sources, etc., can solve problems such as unsatisfactory prognosis, and achieve the effect of preventing and treating tumor cell metastasis

Inactive Publication Date: 2006-11-29
PEKING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the treatment of lung cancer is mainly adjuvant treatment methods such as traditional surgical resection and radiotherapy and chemotherapy. However, most patients have entered the middle and late stages when they are diagnosed and have already metastasized when they are discovered, so the prognosis is still not ideal.

Method used

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  • Tumour antigen protein and tumour antigen peptide

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1. Discovery of BJ-TSA-9 full-length cDNA in tumor tissue

[0027] According to the data in the Unigene and EST databases in Genbank, using online bioinformatics tools such as SAGE and virtualNortherns, the sequence on chromosome 8 was analyzed, and it was found that the 24th region of the long arm of chromosome 8 may encode a tumor-specific gene. Therefore, specific PCR primers were designed according to its sequence, and the full-length mRNA sequence of BJ-TSA-9 was successfully cloned from lung cancer tissue.

[0028] Firstly, total RNA was extracted from lung cancer and adjacent tissues with TRIzol kit (Gibco Company), and the concentration and purity of mRNA were detected by ultraviolet spectrophotometry. Reverse transcription was performed using the advantage reverse transcriptase kit (Clontech Company), and cDNA was synthesized from 2 μg of total RNA. After diluting to 100ul, take 2.5ul as a template, and perform PCR with high-fidelity pfu DNA synthetase...

Embodiment 2

[0028] Firstly, total RNA was extracted from lung cancer and adjacent tissues with TRIzol kit (Gibco Company), and the concentration and purity of mRNA were detected by ultraviolet spectrophotometry. Reverse transcription was performed using the advantage reverse transcriptase kit (Clontech Company), and cDNA was synthesized from 2 μg of total RNA. After diluting to 100ul, take 2.5ul as a template, and perform PCR with high-fidelity pfu DNA synthetase, the conditions are 94°C for 20 seconds; 60°C for 30 seconds; 72°C for 1 minute and 30 seconds, 35 cycles, and 72°C for 7 minutes . The primer sequence is forward: 5'-CAC TTC TTG GAG GTG CCC TGCACG-3'; reverse: 5'-CTC CCA AGC CCA GCT TTC CAA AGG-3'. 8 μl of the PCR product was analyzed by 1% agarose gel electrophoresis, and after using Taq DNA polymerase to add a polyA tail, it was cloned into pGEM-T Easy vector according to the conventional method, transformed into competent E.coli TOP10F' bacteria, and sent to Haiji Kang comp...

Embodiment 3

[0031] Embodiment 3, Northern blot hybridization to analyze the expression of BJ-TSA-9 gene in lung cancer tumor tissue

[0032] Use TRIZOL reagent (PROMEGA) to extract total RNA from the cancer tissue and paired paracancerous tissues of lung cancer patients, denature 20ug of total RNA in the presence of formamide and formaldehyde, perform agarose electrophoresis, and transfer to Hybond-N+nylon membrane (Amersham) and fixed by UV crosslinking. The DNA fragments amplified in Example 2 were labeled with Digoxigenin DNA Probe Labeling Kit (ROCHE) to make DNA probes, hybridized with the tRNA on the membrane according to the conventional Northern hybridization method, and then carried out autoradiography, The mRNA of the tumor-specific antigen protein gene BJ-TSA-9 in the present invention was detected. Then, boil the membrane used to detect the gene mRNA in 1% SDS solution, remove the digoxin-labeled probe, use the labeled housekeeping gene G3PDH as a probe, and use the same meth...

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Abstract

A tumor antigen protein and its tumor antigen peptide used to prepare the medicine for treating lung cancer are disclosed. Said tumor antigen protein can generate the peptide fragment by cell endolysis. Said peptide fragment can bind with the (MHC)-I molecular compatible with main tissue and can be recognized by T cells. The DNA sequence coding said tumor antigen protein has the nucleotide sequence No.AF497803 in GenBank.

Description

technical field [0001] The invention relates to a tumor-specific antigen protein and tumor antigen peptide, in particular to a tumor antigen protein and tumor antigen peptide for treating lung cancer. Background technique [0002] As a new mode of tumor treatment, tumor biotherapy has gradually become an irreplaceable and important part of comprehensive tumor treatment, which mainly includes two aspects of immunotherapy and gene therapy. Based on the analysis of current international research trends and from the perspective of practical application, it is speculated that tumor immunotherapy will have broad application prospects. The traditional method is to vaccinate patients with tumor cells, tumor extracts or certain components as antigens, and achieve the purpose of controlling tumors by stimulating immune responses. However, in the absence of identifiable tumor-specific antigens, the elicited nonspecific immune activity is limited in controlling tumor growth. [0003] ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/435C07K14/47C12N15/12A61K38/17A61K39/00A61P35/00
Inventor 陈慰峰李云燕尹艳慧董学员苏艳荣
Owner PEKING UNIV
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