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Method for preparing genotype F mumps attenuated live vaccine

An attenuated live vaccine, mumps technology, applied in the direction of microorganism-based methods, biochemical equipment and methods, inactivation/attenuation, etc., can solve problems such as allergic reactions, affecting product stability, and differences in virus sensitivity, and achieve The effect of good security

Inactive Publication Date: 2014-04-30
BEIJING MINHAI BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

As a heterogeneous cell matrix, chicken embryo cells (CEC) may cause allergic reactions; may carry infectious avian retroviruses; and the quality of each batch of primary cells is unstable, resulting in differences in sensitivity to the same virus, affecting Insufficient product stability

Method used

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  • Method for preparing genotype F mumps attenuated live vaccine
  • Method for preparing genotype F mumps attenuated live vaccine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1F genotype mumps live attenuated vaccine and its preparation method

[0026] Take the human embryonic lung diploid cells MHL-3 (MHL-3 purchased from Beijing Minhai Biotechnology Co., Ltd.) that grows into a single layer, and digest them with 0.25% trypsin solution. After the cells are soaked in trypsin solution, they form single cells. , Pour out the trypsin solution in the cell flask, add MEM cell culture solution pre-warmed at 37℃ and dilute to 10 5 Cells / mL. Then inoculate it in the cell factory and place it at 37°C for 24-48h.

[0027] When the MHL-3 cells grow into a monolayer, discard the original culture medium, and wash the cell surface and the side wall of the cell factory with Earl's solution. F-genotype mumps virus MHM-19 suspension was inoculated at 0.1 MOI, and MEM maintenance solution was added to culture at 33°C.

[0028] Among them, the MEM culture medium contains 0.03% glutamine, 0.08% sodium bicarbonate, and 10% calf serum; the MEM maintenance medi...

Embodiment 2

[0037] Example 2 Quality Inspection of F Genotype Mumps Live Attenuated Vaccine

[0038] The finished product of F genotype mumps live attenuated vaccine prepared in Example 1 was tested. The test items include: identification test, appearance inspection, moisture test, virus titer, heat stability test, bovine serum albumin residue test, sterility test, and abnormal toxicity test. The test results are shown in Table 2.

[0039] Table 2 F genotype mumps live attenuated live vaccine related test results

[0040]

Embodiment 3

[0041] Example 3 Immunogenicity analysis of F genotype mumps live attenuated vaccine

[0042] Fifteen rhesus monkeys (age 2-3 years old, weight 3kg±0.5kg) negative for mumps antibody were screened, and then divided into 3 groups, MHM-19 strain (prepared in Example 1) and S79 mumps attenuated live The vaccines were used to immunize a group of rhesus monkeys, and the negative control group was injected with 0.5ml DMEM. The immunization method was subcutaneous. At 2, 4, 8, 12, and 24 weeks after immunization, blood was collected to separate serum for neutralizing antibody determination. The results are shown in Table 3. The serum neutralizing antibody titers of the negative control group were all less than 2, which was negative. Both the MHM-19 and S79 experimental groups had good neutralizing antibody responses, and the immune effects of MHM-19 and S79 strains Significantly different (P <0.05). During the test period, the rhesus monkeys in each test group had no signs of parotid ...

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Abstract

The invention provides a method for preparing genotype F mumps attenuated live vaccine. The method comprises the following steps of inoculating mumps attenuated strain MHM-19 to human embryo lung diploid cell MHL-3 which is cultured to a monolayer; releasing virus through frozen-thawed cell in the peak of viral multiplication; centrifuging and removing cell debris; adding a protective agent to centrifuged virus suspension to obtain a semi-finished product of vaccine; and freezing and drying the semi-finished product to obtain the finished product of genotype F mumps attenuated live vaccine. After a rhesus monkey is injected with the prepared MHM-19 attenuated live vaccine for immunizing, good humoral immunity is induced to be produced, and the detection results obtained at different times after immunization show that the MHM-19 attenuated live vaccine has immunizing potency superior to that of reference mumps vaccine S79; in addition, the safety experiment shows that the rhesus monkey does not suffer from parotitis-related symptom after being injected with MHM-19 attenuated live vaccine, the pathological tissue nearly has no obvious difference with that of S79 strain by observing, so that the MHM-19 attenuated live vaccine has high safety.

Description

Technical field [0001] The invention relates to the field of vaccine preparation, in particular to a method for preparing an F genotype mumps live attenuated vaccine. Background technique [0002] Mumps, referred to as mumps or mumps, is an acute systemic infectious disease caused by the mumps virus. Mumps is prevalent all over the world, and it can occur throughout the year, mainly in winter and spring. The disease can occur at any age, most patients are 5-15 years old, and the disease rarely occurs under 1 year old and over 15 years old. After infection, lifelong immunity is generally available. The incubation period of infection is 14-25 days, with an average of 18 days. The clinical feature is parotid gland swelling and pain, which may be accompanied by fever and headache. Early patients and recessive infections are sources of infection. Spread through air droplets or direct contact with saliva and saliva-contaminated food utensils and toys. MuV is the most common cause...

Claims

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Application Information

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IPC IPC(8): A61K39/165A61P31/14C12N7/08C12R1/93
CPCA61K39/12A61K2039/5254A61P31/14C12N2760/18734
Inventor 柳洪涛李黎张现臣刘雨孟红彦黄秋香魏文进
Owner BEIJING MINHAI BIOTECH
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