Unlock instant, AI-driven research and patent intelligence for your innovation.

Application of Flow Cytometry in Real-time Monitoring of Vaccine Production

A technology of flow cytometry and vaccine production, which is applied in the application field of flow cytometry method in real-time monitoring of vaccine production, can solve the problems of inability to control the fermentation process of bacterial liquid in real time, high sensitivity of culture method, long time consumption, etc. Achieve the effect of reliable bacterial activity monitoring method and precise quality control

Active Publication Date: 2016-08-24
INST OF ANIMAL HEALTH GUANGDONG ACADEMY OF AGRI SCI
View PDF4 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The culture method has high sensitivity but takes a long time, and it usually takes more than 24 hours to obtain the result, which cannot achieve the purpose of immediate control of the fermentation process of the bacterial liquid; the turbidimetric method is simple to operate but uses naked eyes to observe, the detection sensitivity is poor, and it cannot distinguish between live bacteria and dead bacteria

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Application of Flow Cytometry in Real-time Monitoring of Vaccine Production
  • Application of Flow Cytometry in Real-time Monitoring of Vaccine Production
  • Application of Flow Cytometry in Real-time Monitoring of Vaccine Production

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0060] (1) Establish the blank control and fluorescence intensity compensation control of Mycoplasma gallisepticum cells

[0061] ① Activate Mycoplasma gallisepticum F-36 strain (China Veterinary Drug Administration) with modified Frey's medium, culture at 37°C for more than 80 hours, inoculate shake flask fermentation medium (improved Frey's medium), and culture at 37°C until bacterial growth In the logarithmic phase, take 2 tubes of fermentation broth, 1 mL each; centrifuge the fermentation broth (9000 rpm for 10 min), discard the supernatant, and obtain bacterial cells. Add 1 mL of ethanol solution with a mass volume ratio of 70% to one of the tubes of bacterial cells, mix well and let it stand for 1 hour, or add 1 mL of PBS, and boil for 10 minutes, the purpose is to inactivate and obtain the dead bacteria control; Treated bacterial cells were used as viable control.

[0062] ② Wash the two kinds of cells in step ① twice with PBS (pH7.4, 0.01mol / L), and then resuspend the...

Embodiment 2

[0078] (1) Establish the fluorescence intensity blank control and compensation control of Clostridium welchii cells

[0079]① Clostridium welchii (ATCC13124, purchased from American type culture collection) was inoculated with liquid thioglycollate (FT) medium for seed medium, activated at 37°C for 24 hours, and inoculated with shake flask fermentation medium (FT culture medium). base), cultured at 37°C for 24 hours to the logarithmic phase of bacterial growth, took 2 tubes of fermentation broth, 1 mL each; centrifuged the fermentation broth (5000 rpm for 10 min), discarded the supernatant, and obtained bacterial cells. Add 1mL PBS (pH6.2, 0.01mol / L) to one tube of bacterial cells, boil for 5min, the purpose is to inactivate and obtain dead bacteria control; at the same time, use the other tube of untreated bacterial cells as a live bacteria control .

[0080] ②Wash the two kinds of cells in step ① twice with PBS, and then resuspend to OD with PBS 600 is 0.004.

[0081] A. ...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

PropertyMeasurementUnit
particle diameteraaaaaaaaaa
Login to View More

Abstract

The invention discloses the application of a flow cytometry method in real-time monitoring of vaccine production. The application is to apply the flow cytometry method to the vaccine production process to monitor the number of live bacteria in the vaccine production process in real time. The present invention stains bacterial cells with fluorescent dyes, detects changes in bacterial fluorescence signals with a flow cytometer, and obtains FCM spectra with at least two of the following parameters by measuring the changes in bacterial fluorescence information: the number of viable bacteria, the number of dead The number of bacteria and the total number of bacteria; the ratio of the number of viable bacteria obtained according to the FCM map; the concentration of the target bacteria solution obtained by the absolute counting method; the number of viable bacteria of the target bacteria was obtained according to the ratio of the number of viable bacteria and the concentration of the target bacteria solution and dead bacteria. The invention provides a simple, rapid and reliable bacterial activity monitoring method for the vaccine production process, which is simple and easy to operate, only needs 30 minutes of operation time, and can accurately control the quality of the vaccine production process.

Description

technical field [0001] The invention relates to the technical application of flow cytometry, in particular to the application of the flow cytometry method in real-time monitoring of vaccine production. Background technique [0002] The control of the bacterial fermentation process in the vaccine production process is a key link in vaccine production. Various factors such as temperature, pH value, and nutritional status of the culture medium all affect the fermentation process of the bacterial solution. Therefore, the establishment of a simple, rapid and reliable detection method for bacterial activity has practical significance for the monitoring of vaccine production. Traditional methods for monitoring vaccine production mainly include culture counting and turbidimetric counting. At present, vaccine manufacturers still rely on these two methods for vaccine quality control. The culture method has high sensitivity but takes a long time. It usually takes more than 24 hours t...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Patents(China)
IPC IPC(8): G01N15/14
Inventor 孙俊颖刘志成郭鹏举
Owner INST OF ANIMAL HEALTH GUANGDONG ACADEMY OF AGRI SCI