A preparation method and application of liver cancer immunogen z1

An immunogen, liver cancer technology, applied in the field of preparation of liver cancer immunogen Z1, can solve the problems of liver cancer prevention blank, immune system to prevent liver cancer, etc., to achieve the effect of protecting immune organs, prolonging survival rate, and improving cellular immunity

Active Publication Date: 2017-10-27
潍坊峡山维泰生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

So far, the research on liver cancer has only achieved certain results in the treatment, but there is still a gap in the prevention of liver cancer. The immunotherapy method currently used is only applied to postoperative recovery, and does not really use the immune system to recover from the disease. From the perspective of prevention, prevent the occurrence of liver cancer

Method used

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  • A preparation method and application of liver cancer immunogen z1
  • A preparation method and application of liver cancer immunogen z1
  • A preparation method and application of liver cancer immunogen z1

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Experimental program
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Embodiment 1

[0030] This embodiment provides a method for preparing liver cancer immunogen, the steps are as follows:

[0031] 1) In 4 mL of RPMI1640 culture medium containing 10% inactivated fetal bovine serum, inoculate mouse H22 liver cancer cells in the logarithmic growth phase to a cell concentration of 2-6.25×10 7 per mL, add 1 mL of 175 μg / mL lentinan at the same time, at 37°C, CO 2 Cultivate for 48 hours under the condition of a concentration of 5%, and obtain the culture after the end of the cultivation;

[0032] 2) Using liquid nitrogen to repeatedly freeze and thaw the culture obtained in step 1) 5 times, centrifuge at 4°C and 4000r / min for 5min to obtain the supernatant;

[0033] 3) Use ammonium sulfate with 30% saturation to stir at 4°C to process the supernatant obtained in step 2) for 60min, then stand still at 4°C for 1.5h, centrifuge at 4°C, 10000r / min for 5min, and collect the precipitate , the liver cancer immunogen Z1 was obtained by using a dialysis membrane with a m...

Embodiment 2

[0035] This embodiment provides a method for preparing liver cancer immunogen, the steps are as follows:

[0036] 1) In 4 mL of RPMI1640 culture medium containing 10% inactivated fetal bovine serum, inoculate mouse H22 liver cancer cells in the logarithmic growth phase to a cell concentration of 2-6.25×10 7 per mL, add 1 mL of 50 μg / mL lentinan at the same time, at 37°C, CO 2 Cultivate for 48 hours under the condition of a concentration of 5%, and obtain the culture after the end of the cultivation;

[0037] 2) Using liquid nitrogen to repeatedly freeze and thaw the culture obtained in step 1) 5 times, centrifuge at 4°C and 4000r / min for 5min to obtain the supernatant;

[0038] 3) Use ammonium sulfate with 30% saturation to stir at 4°C to process the supernatant obtained in step 2) for 60min, then stand still at 4°C for 1.5h, centrifuge at 4°C, 10000r / min for 5min, and collect the precipitate , the liver cancer immunogen Z1 was obtained by using a dialysis membrane with a mo...

Embodiment 3

[0040] This embodiment provides a method for preparing liver cancer immunogen, the steps are as follows:

[0041] 1) In 4 mL of RPMI1640 culture medium containing 10% inactivated fetal bovine serum, inoculate mouse H22 liver cancer cells in the logarithmic growth phase to a cell concentration of 2-6.25×10 7 per mL, add 1 mL of 100 μg / mL lentinan at the same time, at 37°C, CO 2 Cultivate for 48 hours under the condition of a concentration of 5%, and obtain the culture after the end of the cultivation;

[0042] 2) Using liquid nitrogen to repeatedly freeze and thaw the culture obtained in step 1) 5 times, centrifuge at 4°C and 4000r / min for 5min to obtain the supernatant;

[0043] 3) Use ammonium sulfate with 30% saturation to stir at 4°C to process the supernatant obtained in step 2) for 60min, then stand still at 4°C for 1.5h, centrifuge at 4°C, 10000r / min for 5min, and collect the precipitate , the liver cancer immunogen Z1 was obtained by using a dialysis membrane with a m...

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Abstract

The invention discloses a preparation method and application of liver cancer immunogen Z1, belonging to the technical field of bioengineering. The method provided by the present invention is to inoculate H22 liver cancer cells in the culture medium and add lentinan for co-cultivation, take out after the culture, repeatedly freeze-thaw and break the cells, centrifuge to obtain the supernatant, add ammonium sulfate to the supernatant for salting After the salting out, the precipitate was separated and collected, and finally the liver cancer immunogen Z1 was obtained. Utilizing the immunogen prepared by the preparation method provided by the present invention to inoculate mice can effectively prolong the survival rate of mice. Compared with the control group, the survival rate of mice inoculated with the immunogen prepared by the present invention can be increased by 100%. The index and spleen index reached 2.68 and 4.56 respectively, the phagocytosis rate and phagocytosis index of macrophages reached 52% and 3 respectively, and the mouse serum antibody titer reached 25600×.

Description

technical field [0001] The invention relates to a preparation method and application of liver cancer immunogen Z1, belonging to the technical field of bioengineering. Background technique [0002] At present, the number of patients with liver cancer in my country is increasing year by year. Lentinan, as a functional factor with good physiological activity, is of great significance in the treatment and rehabilitation of patients with liver cancer. According to the location of the tumor, liver cancer is divided into two types: primary and secondary. It has the characteristics of long incubation period, high degree of malignancy, rapid progression, easy metastasis, strong invasiveness, insidious onset, and poor prognosis. [0003] Every year, the number of newly discovered malignant tumor patients in the world is about 6.35 million, of which liver cancer accounts for 4% of malignant tumors, that is, 60,000 cases, and the incidence of liver cancer around the world is also increa...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/00A61P35/00
Inventor 韩雪赵诗杨王颖张东杰李研东
Owner 潍坊峡山维泰生物科技有限公司
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