TM4SF3 protein for preparing antineoplastic antibody and use thereof

A protein and antibody technology, which is applied in the field of TM4SF3 protein and its encoded nucleotide sequence for the preparation of anti-tumor antibodies, can solve problems such as cell damage, reduce tumor mortality, and achieve good anti-tumor growth effect.

Inactive Publication Date: 2012-07-04
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

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

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

[0034] According to the prior art in the field, using the cDNA sequence of the TM4SF3 gene as a template, primers were used: 5-ATAGATATCGACAAGCCTGTAACGAA-3' (forward primer, containing EcoRV restriction site and XhoI restriction site) and 5'-GATCTCGAGGTTCCCGATCTGG- 3' (the reverse primer contains the XhoI restriction site), the cDNA coding sequence of the TM4SF3 gene was cloned by PCR technique. After the cloned PCR product was verified to be the correct cDNA sequence of the TM4SF3 gene by sequencing, 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 TM4SF3 protein. ...

Embodiment 2

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

[0039] Through Westen detection, the inventors found that the high-frequency expression of TM4SF3 protein was up-regulated in esophageal cancer tissues.

[0040] Westen detected the expression of TM4SF3 protein in 14 pairs of esophageal cancer (T) and its paired adjacent 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 by SDS-PAGE electrophoresis was 50ug. The results of Westen detection showed that the expression of TM4SF3 in esophageal cancer tissues was up-regulated in 11 cases among all 14 pairs of esophageal cancer and their paired normal esophageal epithelial tissues, and the up-regulation rate reached 78.6% (figure 1 ). Therefore, the TM4SF3 protein is a specific human tumor antigen and can b...

Embodiment 3

[0041] Example 3: Preparation of new human tumor antigen corresponding antibody

[0042] Using 200 μg of human tumor antigen (TM4SF3 protein) obtained in Example 1 or its fragments, 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 of antigenic protein or its fragments after 2 weeks Boost immunization subcutaneously with incomplete Freund's adjuvant. One week later, use complete Freund's adjuvant to emulsify 50-60 mg BCG protein and inject bilateral paws. One week later, use 200 μg of antigenic protein or its fragments to mix equal volumes of incomplete Freund's adjuvant. 200ul was injected into the popliteal lymph nodes on both sides, blood was collected 10 days later and the titer was measured by conventional methods, and the titer of the serum was found to be >1:10 8 , sacrifice the animals, collect rabbit serum, and use methods known to those skilled in the art, ...

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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 thatthe 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 preparing anti-tumor antibodies; the anti-TM4SF3 protein antibody and its preparation method, and its use in the production of various biological agents for the treatment of various related tumors; in addition 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 human diseases, and its death rate ranks second and third among various diseases in the world and our country, respectively, seriously affecting human health. With the development of society in our country, the incidence of tumor is still increasing year by year. There are currently about 4.5 million ...

Claims

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Application Information

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Patent Type & Authority Patents(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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