Preparation method and application of liver cancer immunogen Z1

An immunogen and liver cancer technology, applied in the field of preparation of liver cancer immunogen Z1, can solve the problems of immune system preventing liver cancer, liver cancer prevention blanks, etc., and achieve the effects of protecting immune organs, prolonging survival rate, and improving humoral immunity.

Active Publication Date: 2015-06-24
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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  • Preparation method and application of liver cancer immunogen Z1
  • Preparation method and application of liver cancer immunogen Z1
  • Preparation method and application of liver cancer immunogen Z1

Examples

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 biological engineering. The preparation method provided by the invention comprises the following steps of: inoculating H22 liver cancer cells in a culture medium, simultaneously adding lentinan to co-culture, taking out after culturing, repetitively freezing, thawing and crushing the cells, centrifugally separating to obtain supernate, adding ammonium sulphate into the supernate to salt out, and separating and collecting precipitate after salting out so as to finally obtain the liver cancer immunogen Z1. If a mouse is inoculated with the immunogen prepared by the preparation method provided by the invention, the survival rate of the mouse can be effectively prolonged; compared with a control group, the survival rate of the mouse inoculated with the immunogen prepared by the invention can be increased by 100%; the thymus index and the spleen index are up to 2.68 and 4.56 respectively; the phagocytic rate and the phagocytic index of macrophage are up to 52% and 3 respectively; and the serum antibody titer of the mouse is up to 25600x.

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 Applications(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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