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Recombination immune therapeutic protein and its expression method and application

A technology of immunotherapy and expression method, which is applied in the field of genetic engineering to achieve the effect of reducing requirements, alleviating anti-rejection reactions, and promoting wound healing

Inactive Publication Date: 2006-11-29
THE SECOND AFFILIATED HOSPITAL OF GUANGZHOU MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] The purpose of the present invention is to provide a recombinant immunotherapeutic protein to overcome the obstacles in the application of a large number of recombinant allergens or extracts for the problems existing in the existing immunotherapy, and to apply it to allergic atopic people and other types of immunotherapeutics. diseases, to improve the efficacy of immunotherapy and shorten the treatment cycle, thereby replacing allergen immunotherapy

Method used

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  • Recombination immune therapeutic protein and its expression method and application
  • Recombination immune therapeutic protein and its expression method and application
  • Recombination immune therapeutic protein and its expression method and application

Examples

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example 1

[0040] Example 1 Expression and purification of recombinant broad-spectrum immunotherapeutic protein

[0041] (1) Obtain the target gene: extract the total RNA of ragweed pollen, perform polymerase chain reaction after reverse transcription, recover the band obtained by PCR and connect it to the pGEM-T vector, transform Escherichia coli DH5a, and the positive clone is confirmed by sequencing Finally, the full-length allergen gene TT is obtained; primers are designed according to the obtained gene sequence, and the cDNA ends are rapidly amplified to obtain different cDNA ends, and the new pan-allergen full-length gene TT that has been modified by bridge PCR is obtained. ; PCR annealing temperature is 58 ~ 68 ℃.

[0042] (2) Collect human peripheral blood, use gradient centrifugation Ficoll method to separate mononuclear cells and extract RNA; use RT-PCR method to amplify the interleukin full-length cDNA, parallel T / A clone, after sequencing verification, use bridge The new int...

example 2

[0047] Example 2 The therapeutic effect of recombinant broad-spectrum immunotherapeutic protein on scald

[0048] Dissolve the purified target protein in PBS solution, prepare three concentrations of recombinant broad-spectrum immunotherapeutic protein (64 μg / ml, 320 μg / ml, 1600 μg / ml), and use commercially available Scald No. 1 and Scald No. 2 as controls , to conduct treatment experiments on scald model animals. First, establish a scalded animal model: randomly select 10 adult rats of the right age, male and female, weighing more than 250 g, and number them. After 1 week of routine feeding, remove the long hair on the back of the rat with a depilatory agent, with an area of ​​about 1.5 × 1.5 cm. , entered the test after feeding for 1 day. Cut out 1.5 x 1.5 cm square pieces of gauze, soak in boiling water for later use. After depilatory rats were anesthetized with ether, quickly stick the gauze soaked in boiling water on the depilatory area, start timing, remove the gauze a...

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Abstract

A recombinant protein used for immunotherapy by regulating the balance between Th1 and Th2 in human body is composed of the variant derived from sensibiligen and the variant derived from interleukin. It is prepared through cloning the pan-sensibiligen of pollen and full-length coding gene of interleukin, taking their open reading frames, antigen modifying, gomphosis, and carrying it by protein expression carrier.

Description

technical field [0001] The invention relates to the field of genetic engineering, in particular to a recombinant immunotherapeutic protein and its expression method and application. Background technique [0002] Allergic diseases are increasing with the advancement of urbanization and industrialization, becoming an increasingly prominent health problem, which has attracted widespread attention of researchers worldwide. During the four years from January 1, 2000 to September 1, 2004, there were more than 30,000 research papers on allergies, accounting for nearly a quarter of the research papers on immunology during the same period . In our country, research on allergens started late, especially research on allergen gene cloning, allergen standardization, and hypoallergenic allergen (Hypoallergen) vaccines was relatively weak. In terms of diagnosis and treatment, allergen extracts are generally used in China, which is contrary to the requirements of modern medicine and may b...

Claims

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Application Information

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IPC IPC(8): C07K19/00C12N15/62C12N15/63C12N15/09A61K38/16A61P17/02A61P29/00
Inventor 陶爱林
Owner THE SECOND AFFILIATED HOSPITAL OF GUANGZHOU MEDICAL UNIV
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