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TM4SF3 protein for preparing antineoplastic antibody and use thereof

A protein and antibody technology, which is applied in the field of preparing anti-tumor antibody TM4SF3 protein and its coding nucleotide sequence, can solve problems such as cell damage and reduce tumor mortality, and achieve good anti-tumor growth effects

Inactive Publication Date: 2009-08-05
CANCER INST & HOSPITAL CHINESE ACADEMY OF MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The main methods of clinical treatment of cancer are surgical resection and radiotherapy and chemotherapy. However, while radiotherapy and chemotherapy kill cancer cells, they also cause serious damage to normal cells in the human body.
The three conventional treatment methods of surgery, radiotherapy and chemotherapy for the treatment of malignant tumors have not significantly reduced the mortality rate of tumors, and their 5-year survival rate is only 10-30%. Therefore, countries all over the world are constantly exploring and trying to find more effective treatments

Method used

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  • TM4SF3 protein for preparing antineoplastic antibody and use thereof
  • TM4SF3 protein for preparing antineoplastic antibody and use thereof
  • TM4SF3 protein for preparing antineoplastic antibody and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1: Preparation of new human tumor antigen

[0034] According to the prior art in the field, using the cDNA sequence of the TM4SF3 gene as a template, primers: 5'-ATAGATATCGACAAGCCTGTAACGAA-3' (forward primer containing EcoRV restriction site and XhoI restriction site) and 5'-GATCTCGAGGTTCCCGATCTGG -3'(Reverse primer contains XhoI restriction site), clone the cDNA coding sequence of TM4SF3 gene by PCR technology. After sequencing and verifying the cloned PCR product to be the correct cDNA sequence of the TM4SF3 gene, the sequence was constructed in the expression vector PET-30b according to conventional techniques in the art to obtain the recombinant vector PET-30b-M4SF3. Then the recombinant vector PET-30b-TM4SF is transformed into prokaryotic cells to obtain corresponding recombinant prokaryotic cells, and the TM4SF3 gene is expressed under conditions suitable for the expression of the recombinant vector to obtain the TM4SF3 protein.

[0035] The amino acid sequenc...

Embodiment 2

[0038] Example 2: TM4SF3 is highly expressed in esophageal cancer tissue

[0039] Through Westen detection, the inventors found that TM4SF3 protein is highly expressed in esophageal cancer tissues.

[0040] Westen detected the expression of TM4SF3 protein in 14 pairs of esophageal cancer (T) and its paired normal esophageal epithelial tissue specimens (N). The total protein of each tissue was extracted, and the total protein of each tissue obtained was detected by SDS-PAGE electrophoresis and Westen detection. The total protein of each tissue sample loaded on SDS-PAGE electrophoresis was 50ug. Westen test results showed that TM4SF3 expression was up-regulated in 11 cases of esophageal cancer tissues in all 14 pairs of esophageal cancers and their paired adjacent normal esophageal epithelial tissues, with an up-regulation rate of 78.6% (figure 1 ). Therefore, TM4SF3 protein is a specific human tumor antigen and can be used in the development of further preparation of anti-tumor an...

Embodiment 3

[0041] Example 3: Preparation of new antibodies corresponding to human tumor antigens

[0042] Using 200 μg of human tumor antigen (TM4SF3 protein) or fragments obtained in Example 1, mixed with an equal volume of complete Freund’s adjuvant for a total of 200 ul, immunized New Zealand rabbits by subcutaneous immunization, and mixed with 200 μg antigen protein or fragments after 2 weeks Incomplete Freund’s adjuvant is subcutaneously boosted. After 1 week, 50-60mg of BCG protein is emulsified with complete Freund’s adjuvant and injected into the soles of both sides. After 1 week, 200μg of antigen protein or its fragments are mixed with equal volume of incomplete Freund’s adjuvant. 200ul was injected into the popliteal lymph nodes on both sides. After 10 days, blood was collected and the titer was measured by conventional methods. It was found that the titer of the serum was> 1:10 8 The animals were sacrificed, and rabbit serum was collected, using methods known to those skilled in t...

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Abstract

The invention relates to a TM4SF3 protein used for preparing an anti-tumor antibody, and an application thereof, in particular to a TM4SF3 protein used for preparing an anti-tumor antibody, and an encoded nucleotide sequence thereof; the invention also relates to an antibody of the anti-TM4SF3 protein, and a preparation method and an application of the antibody in producing various biological preparations for curing various relative tumors; in addition, the invention also relates to a medicine composition including the antibody, and an application for inhibiting the tumor in the body, so that the effect of curing the tumor is achieved.

Description

Technical field [0001] The present invention relates to TM4SF3 protein which can be used for preparing anti-tumor antibody and its application. Specifically, the present invention relates to the TM4SF3 protein and its encoding nucleotide sequence for the preparation of anti-tumor antibodies; relates to the anti-TM4SF3 protein antibody and its preparation method, and its use in the production of various biological preparations for the treatment of various related tumors; It also relates to a pharmaceutical composition comprising the antibody and its use in inhibiting tumors in vivo, so as to achieve the effect of treating tumors. Background technique [0002] Tumor is one of the most common diseases in humans, and its mortality rate ranks second and third in various diseases in the world and my country, which seriously affects human health. With the development of society in our country, the incidence of tumors is still increasing year by year. There are currently about 4.5 millio...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/705C07K16/30A61K39/395A61P35/00G01N33/68
Inventor 杨治华冉宇靓胡海
Owner CANCER INST & HOSPITAL CHINESE ACADEMY OF MEDICAL SCI
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