Tumor individualized specific therapeutic vaccine and preparation method thereof

A technology for therapeutic vaccines and tumor patients, applied in the direction of anti-tumor drugs, antibody medical ingredients, medical preparations containing active ingredients, etc., can solve the problem of efficient expansion and maturation of DC cells, difficult induction of CTL, and patients Long waiting time and other problems, to achieve the effect of shortening the waiting time, outstanding treatment effect, and short treatment cycle

Pending Publication Date: 2020-08-04
河北博海生物工程开发有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The biggest problem with these antigens is that there is no specific screening, that is, there is no specific screening on normal tissue banks and tumor tissue banks. Most of them have cross-reactivity and poor specificity. It is difficult to make DC cells highly efficient in vitro. Expand and mature, it is difficult to induce specific CTL
[0014] (3) Long preparation period
This treatment plan has high specificity, but it takes a long time for gene sequencing and gene translation to synthesize protein. It takes about 1-2 months from the beginning of detection to the cultivation of qualified DC cells. In clinical application, the patient waiting time longer

Method used

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  • Tumor individualized specific therapeutic vaccine and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] Example 1: Molecular target detection, analysis and synthesis

[0051] 1. Sample collection: collect 5ml of blood sample to be tested according to clinical blood collection requirements, separate serum, and keep at 2-8°C

[0052] Transportation, ensure delivery to the laboratory within 72 hours.

[0053] 2. Virus detection and sample retention:

[0054] 3. Enrichment: add the serum to be tested into phosphoric acid solution and acetonitrile solution, the volume ratio of serum, phosphoric acid and acetonitrile is 1:1-2:4-6; mix well and centrifuge to get the supernatant;

[0055] 4. Purification: add acetonitrile solution to the supernatant and mix evenly, acetonitrile: supernatant volume ratio is 0.3-1:2, centrifuge to get the supernatant and concentrate to 1-50 μl, then add water to dilute;

[0056] 5. Collection: filter through a hydrophobic solid phase extraction column Oasis Prime HLB, wash the solid phase extraction column with methanol, elute with a mixed soluti...

Embodiment 2

[0060] Example 2: In vitro amplification and activation method for tumor-specific dendritic cell targets

[0061] Blood sample collection: Collect 80--120ml of peripheral blood from patients according to clinical blood collection requirements, transport at 2-8°C, and ensure delivery to the laboratory within 6 hours.

[0062] Separation of PBMCs: Mononuclear cells (PBMCs) in blood were separated by density gradient centrifugation with lymphocyte separation medium.

[0063] Culture of DC cells and MCTL cells: According to the results of PBMC counting, adjust the cells to about 1×10 with culture medium 7 cells / ml, inoculate, and incubate in a carbon dioxide incubator at 37°C; 5% CO 2 ; Incubation for 30min.

[0064] Culture the adherent cells after incubation as DC: take out the culture flask, shake gently to float the suspended cells that settled at the bottom, and transfer to a centrifuge tube. After adhering to the wall, wash repeatedly with normal saline several times unti...

Embodiment 3

[0068] Example 3: In vitro amplification and activation method of tumor-specific dendritic cell (DC) target

[0069] 1. Specimen collection:

[0070] The blood picture of melanoma patients is within the normal range (WBC: 4-10×10 9 , LYM%: 20%-40%), the fluctuation does not exceed 5%, the circulation volume of peripheral blood by machine sampling is 1000-4000ml, the number of mononuclear cells: more than 1×10 9 . Blood drawing method 50-100ml, the number of mononuclear cells: more than 1×10 6 .

[0071] 2. Isolation of Mononuclear Cells

[0072] Transfer the PBMC cell suspension sample, pour it into a centrifuge tube and centrifuge, balance the centrifuge tube, centrifuge at 1310g for 10min, RT. The plasma layer was aspirated and discarded. Dilute the above cell pellet with normal saline at a ratio of 1:1 and mix by pipetting. Slowly add 30ml of the diluted cell suspension into a 50ml sterile centrifuge tube containing 15ml of human lymphocyte separation medium at room ...

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Abstract

The invention relates to the field of immunology, and discloses a tumor individualized specific therapeutic vaccine and a preparation method thereof. Through MCTL proteomics dendritic cell cluster target detection, a tumor specific molecular target in a subject is analyzed, a therapeutic target is synthesized in vitro and co-cultured with autoimmune cells of the cultured subject, and the therapeutic vaccine is prepared. After the therapeutic vaccine is transfused back to a subject, tumor cells are specifically killed, immune balance is rebuilt, the purpose of effectively treating tumors is achieved, and the therapeutic vaccine has wide clinical application prospects.

Description

technical field [0001] The invention relates to the field of immunology, in particular to a tumor individualized and specific therapeutic vaccine and a preparation method thereof. Background technique [0002] Malignant tumors are one of the major diseases that seriously affect human health and threaten human life. Its early diagnosis and timely treatment are very important. In the past 20 years, medical diagnostic technology has developed very rapidly, and new diagnostic methods continue to emerge. Not only can tumors as small as 0.5-1.0 cm be found, but also the extent of tumors can be correctly judged. Clinical diagnosis of advanced cancer. [0003] At present, the diagnosis of tumor mainly relies on physical examination, imaging, pathology and related laboratory tests. Physical examination is an important part of tumor diagnosis. On the basis of comprehensive and systematic examination, local examination of key organs is carried out in combination with medical history...

Claims

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Application Information

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IPC IPC(8): A61K39/00A61P35/00
CPCA61K39/0011A61P35/00
Inventor 李彬
Owner 河北博海生物工程开发有限公司
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