This ppt provide information about the conventional methods of animal vaccine production..it is somewhat differ from my earlier ppt of vaccine production techniques..

1. Conventional methods of
animal vaccine production
Submitted by:
Dr. Vijayata
(dr.viz.vet@gmail.com)
1

2. Pathogen
Inactivation Culture
Attenuation
Ag
Purification
Seed (Live
VACCINE Purification attenuated)
Inactivation
wP,
HAV Culture
VACCINE
VACCINE
aP
VACCINE
Rab,Flu
MMR,OPV
Ada G. The immunology of vaccination. In: Plotkin SA, Orenstein WA. Vaccines. 4th
ed. Philadelphia, PA: Saunders, 2003:31-45. Principles of Vaccination
Veterinary Immunology by Tizard

3. Types of Vaccine Manufacturing Methods
Egg Based Vaccine
Production
Cell Culture Based
Vaccine Production
3
Recommendations for production and control of influenza vaccine ,WHO

4. Vaccine production methods
Bacterial Vaccine:
Diseases Vaccines Methods of Production
Dipthera DTaP, DTaP/Hib, Tdap Cell based
Hemophilus infl uenza Hib, DTaP/Hib, Hib/Hepatitis Cell based
type B (Hib)
Pertussis (whooping DTaP, DTaP/Hib, Tdap Cell based
cough)
Pneumococcal Pneumococcal conjugate Cell based
(Heptavalent)
Tetanus DTaP, DTaP/Hib, Tdap Cell based
Meningococcal Meningococcal conjugate Cell based
(quadravalent)
Meningococcal Meningococcal polysaccharide Egg based
(quadravalent)
4
Source : Center for Disease Control and Prevention. VFC: Approved Vaccines and Biologicals. www.cdc.gov/vaccines/programs/vfc/parents/apprvd -
vaccs.htm [accessed September 8, 2007].

5. Viral Vaccine:
Diseases Vaccines Methods of Production
Hepatitis A Hepatitis A (pediatric) Cell based
Hepatitis B Hepatitis B (pediatric/adolescent) Cell based
Hepatitis B Hib/Hepatitis B Cell based
Poliomyelitis (polio) IPV (Inactivated poliovirus vaccine) Cell based
OPV (Oral polio vaccine)
Rotavirus Rotavirus Cell based
Influenza (Flu) Influenza Egg based
Measles MMR, measles Egg based
Mumps MMR, Mumps Egg based
Rubella (German measles) MMR, rubella Egg based
Varicella (chicken pox) Varicella Egg based
5
Source : Center for Disease Control and Prevention. VFC: Approved Vaccines and Biologicals. www.cdc.gov/vaccines/programs/vfc/parents/apprvd -
vaccs.htm [accessed September 8, 2007].

6. Steps in vaccine production
Upstream Downstream
processing processing
Selecting
Harvesti
the Growing Inactivati
ng & Quality
strains the on and Formulat
purificati control
for micro- splitting ion of
on of and lot
vaccine organism of vaccine
microorg release
productio s organism
anisms
n
6
Novel Techniques in the Production of Industrially Imperative Products, Sameera V* Biotechnology Department, REVA Institute of Science and Technology, Bangalore

7. SELECTING THE STRAINS FOR VACCINE
PRODUCTION
• Manufacturing begins with small amounts of a specific
Virus/Bacteria (seed).
• Viruses/Bacteria used in manufacture shall be derived
from a Seed Lot System.
• The virus/Bacteria must be free of impurities, including
other similar viruses/ bacteria and even variations of the
same type of virus/Bacteria.
• The seed shall pass the tests for sterility and freedom
from mycoplasma.
7
Eurropean medical agency guide line for General requirements for the production and control of live mammalian bacterial and viral vaccines for veterinary use (7BIm1a)

8. • A record of the origin, passage history (including purification
and characterisation procedures) and storage conditions should
be maintained for each Seed Lot.
• The seed must be kept under "ideal" conditions, usually frozen,
that prevent the virus from becoming either stronger or weaker
than desired.
Selecting the seed (Strain) used for vaccine production
• The choice of strain depends on a number of factors including
the efficacy of the resulting vaccine, and its secondary effects.
• If possible, the bacterial strain or cell line should be obtained
from a recognized culture collection with an established and
documented provenance.
8
The annual production cycle for influenza vaccine:Catherine Gerdil∗Vaccine 21 (2003) 1776–1779

9. • Alternatively, if the chosen vaccine strain is an “in house”
clinical isolate, it will be necessary to compile a complete history
of the strain, including details of its isolation, identification, and
maintenance for product registration.
Standardizing the seed strains for vaccine production
• Once the candidate seed strains for vaccine production have been
prepared, their antigens undergo identity testings and to evaluate
their suitability for vaccine production.
• This includes satisfactory yields when grown in embryonated
eggs/ cell culture and antigenic stability throughout serial
passage in eggs as well as the inactivation and purification
processes. 9
The annual production cycle for influenza vaccine: Catherine Gerdil∗Vaccine 21 (2003) 1776–1779

10. GROWING THE MICROORGANISMS
• Once the production strain for each vaccine component has been
selected, bulk vaccine production can begin.
• Bulk production begins with the cultivation of the virus or Bacteria
in a fermenter equipped with numerous process parameters to
control temperature, pH, dissolved oxygen, and other factors.
Growing • Batch culture
Growing the Bacteria • Continuous culture
microorganisms
in maximum
titre are mainly
classified in
two category : Growing • Cell (tissue) cultures
The BRIDGE,Cell-Culture-Based Viruses • Embryonated Eggs 10
Vaccine Production: Rino Rappuoli
p 26-27,

11. • Bacteria are grown in
bioreactors e.g. Haemophilus
influenza type b.
Growing
Bacteria • Batch culture
• Continuous culture
Production of Freeze-dried Brucella abortus Strain 19 Vaccine using Cells produced by Continuous Culture† K. J. Boyce, A. W. Edgar, Journal of
Applied Microbiology, Volume 29, Issue 2, pages 401–408, August 1966
11
VACCINE PRODUCTION AS A UNIT PROCESS,PROF.DR.T.O. WIKE'N,1971 Chapter 3.3,pp65-71

12. Batch culture
• The micro-organism are grown in a closed
vessel.
• All the raw materials are put in the fermenter at
the start and then the micro-organism is added.
• The system is then left for a long time –
possibly a week – until all the raw materials
have been used up and there is loads of the
product.
• The fermenter is then emptied and other
processes are used to separate the product from
the micro-organism.
12
VACCINE PRODUCTION AS A UNIT
PROCESS,PROF.DR.T.O.
The tularaemia vaccine: review. Sandstrom, G. Journal of Chemical Technology & Biotechnology.
WIKE'N,1971 Chapter 3.3,pp65-71
Vol. 59, no. 4, pp. 315-20. Apr. 1994

13. Continuous culture
• The micro-organism are grown in an open system.
• Continuous culture aims to keep a culture growing
indefinitely. This can be done if:
• fresh nutrients are continually supplied
• Accumulated cells and waste products are removed at the
same rate
• Conditions such as temperature and pH are kept at their
optimum values.
The tularaemia vaccine: review. Sandstrom, G. Journal of Chemical Technology & Biotechnology. Vol. 59, no. 4, pp. 315-20. Apr. 199413
VACCINE PRODUCTION AS A UNIT PROCESS,PROF.DR.T.O. WIKE'N,1971 Chapter 3.3,pp65-71

14. Here the raw materials are
trickled in at the top of a
column in which there are
immobilised micro-
organisms.
• The product flows out the
bottom in a pure state.
• However this process can
only be used for reactions
that are fast – possibly taking
10 minutes.
• E.g. - in the cultivation of
Corynebacterium diphtheriae
14
VACCINE PRODUCTION AS A UNIT PROCESS,PROF.DR.T.O. WIKE'N,1971 Chapter 3.3,pp65-71

15. • Viruses are grown either on
primary cells e.g.for influenza, or
Growing on continuous cell lines,e.g.
for hepatitis A.
Viruses • Cell culture
• Embryonated Eggs
15
Recommendations for production and control of influenza vaccine ,WHO OIE Terrestrial Manual 2010 C H A P T E R 2 . 8 . 8 . SWINE INFLUENZA,

16. Cell culture
• Cell culture is the complex process by which cells are grown
under controlled conditions, generally outside of their natural
environment.
• Cell cultures are separated into 3 types:-
- Primary cell culture
- Secondary cell culture
- Continuous cell line
16
Mammalian Cell Culture Technology: An Emerging Field ,D. Eibl, R. Eibl, and R. Pörtner

17. Primary cell culture-
• Cells that are cultured directly from animal or human tissues and
can be subcultured only once or twice . e.g. Primary monkey
kidney cell
• For most of the mammalian vaccines the use of primary cells is
not acceptable for the manufacture of vaccines.
• If a vaccine has to be produced on primary cells, they should be
obtained from a SPF herd or flock with complete protection
from introduction of diseases.
Secondary cell culture-
• are derived from human fetal tissue and can be subcultured 20 to
50 times e.g. Human diploid fibroblasts such as MRC-5
17
Mammalian Cell Culture Technology: An Emerging Field ,D. Eibl, R. Eibl, and R. Pörtner

18. Continuous cell lines –
• a single cell type that can propagated indefinitely in culture.
• e.g. Vero cells, Hep2
Cell Line Selection :
• If a cell line is used for the manufacture of vaccines it shall
normally be produced according to a Seed Lot System.
• The history of the cell line must be known in detail (e.g. origin,
number of passages and media used for their multiplication,
storage conditions).
• The cell line used to cultivate the virus must be able to propagate
the virus in large quantities , rapid, efficient in expressing the
desired virus, and suitable for a wide variety of virus strains.
• The cells must be checked for their appearance , rate of growth
and for contamination with bacteria, virus, fungi, mycoplasma.
18
The BRIDGE,Cell-Culture-Based Vaccine Production: Rino Rappuoli p 26-27,

19. • Preparation of a cell line for propagation begins with the
thawing of the cell line seed lot (e.g., EBx™, VERO, or
MDCK).
• Cell line propagation begins with the small scale pre-culture
propagation of seed cells after thawing.
• The cells are then introduced to the fermenter vessel with the
selected nutrient medium.
• When the cell line reaches a predetermined cell density, the
virus is introduced and begins to propagate in the cell line.
• Two methods of mass cultivation of cells are recognized in the
industry today, microcarrier cultures and free-cell
suspension cultures.
• Both systems begin cultivation of the cell line in a fermenter,
which can be scaled up to thousands of liters.
19
The BRIDGE,Cell-Culture-Based Vaccine Production: Rino Rappuoli p 26-27,

20. In microcarrier systems-
• Cells are anchorage dependent and grow on solid or macroporous
microcarriers. (microbeads.)
• Cell expansion often occurs in roller-flasks.
• In the presence of nutrient media, the cells grow and proliferate
covering the beads uniformly.
• Microbeads provide a high surface-area-to-volume ratio, which
can lead to high cell densities.
• Once a bead is covered, the cells are dislodged, dispersed, and
allowed to reattach to achieve another round of cell growth on
the surface of the bead.
20
The BRIDGE,Cell-Culture-Based Vaccine Production: Rino Rappuoli p 26-27,

21. Suspension culture -
• Suspension cultures are derived from cells that can survive and
proliferate without attachment (on-adherent)
• These cells are maintained by continuous stirring with a magnet
and can multiply while suspended in a suitable medium.
• Larger volumes of media are required because the cell line
proliferates while growing freely suspended in the nutrient
medium.
• However, the scaling up of the system is easier, and there is no
limit to the volume.
21
The BRIDGE,Cell-Culture-Based Vaccine Production: Rino Rappuoli p 26-27,

22. Embryonated Eggs
• If the vaccine is to be produced in embryonated eggs, the eggs to
be used should be from closed, specific-pathogen-free, healthy
flocks.
• This flock shall be monitored at regular intervals for Bacteria,
Virus and Mycoplasma.
• Many viruses can be propagated in embryonated chicken eggs
but the method is now only used for Influenza viruses.
• At 5 to 14 days after fertilization, a hole is drilled in the shell
and virus injected into the site appropriate for its replication
(yolk sac, chorioallantoic membrane, amniotic cavity, allantoic
cavity).
• The eggs are incubated at about 33 C for 2 to 3 days, candled
for viability and lack of contamination from the inoculation, and
then the allantoic fluid is harvested.
22
RECOMMENDATIONS FOR PRODUCTION AND CONTROL OF INFLUENZA VACCINE (INACTIVATED) WORLD HEALTH ORGANIZATION)

23. Herpes simplex
virus
Pox virus
Rous sarcoma
virus
Influenza virus
Mumps virus
Influenza virus
Mumps virus
New castle disease
virus
Avian adenovirus
Herpes simplex virus
GROWTH OF VIRUSES IN EMBRYONATED EGG –
An embryonated chicken egg showing the different compartments in which viruses
may grow. The different routes by which viruses are inoculated into eggs are
indicated. 23
http://www.virology.ws/2009/12/10/influenza-virus-growth-in-eggs/

24. Harvesting & Purification of
microorganism
• After propagation, the virus is harvested.
• Harvesting of virus is largely a manual process that requires
extracting infected cells, breaking down cell walls, and then
collecting the virus.
• After treatment of the infected cell line, the virus is released
into the supernatant, and the cellular debris is centrifuged
away by use of appropriate Centrifugation method.
• Purification selectively separates and retains the desired
product at the highest purity per its pre-determined
specification. (Remove unwanted compounds)
24
An international technology platform for influenza vaccines: Jan Hendriksa,∗, Marit Hollemanb, Otto de Boerb, Patrick de Jongc, Willem Luytjesb

25. Centrifugation:
• Centrifugation is a process by which solid particles are
sedimented and separated from a liquid using
centrifugal force as a driving force.
• Centrifugation is used to separation and purification of
pathogenic virus antigens and other agents used in the
production of vaccine.
• Centrifugation is also used to remove dead cells, cell
debris etc.
Example : Influenza vaccine, rabies vaccine , Hepatitis B
vaccine, and Japanese encephalitis vaccine production.
25
An international technology platform for influenza vaccines Filtration Technique in
Jan Hendriksa,∗, Marit Hollemanb, Otto de Boerb, Patrick de Jongc, Willem Luytjesb Vaccine Manufacturing

26. Chromatography
• A group of physical separation techniques, which are
characterized by the separation of mixtures due to differences
in the distribution coefficient of sample components between
two phases, one stationary and the other mobile phase.
• Example : Modified Vaccinia Ankara virus (Small pox
vaccine)
Filtration
• Filtration is a process for separating two substances of two
different physical states. It is used for separating solids from
turbid liquids (filtrate), pure gases or solids.
• Separation of particles from liquid by applying a pressure to the
solution to force the solution through a filter. Filtration Technique in
Vaccine Manufacturing
26
An international technology platform for influenza vaccines
Jan Hendriksa,∗, Marit Hollemanb, Otto de Boerb, Patrick de Jongc, Willem Luytjesb

27. Inactivation And splitting of
microorganism
• Following purification, the virus is inactivated through a
chemical process.
• After inactivation, the whole virus can be purified, split, and ultra
purified as a “subunit.”
• Virus splitting follows because only fractions of specific viral
surface proteins are required for the subsequent vaccine.
(influenza )
• Further purification procedures are then performed.
• At this point, the development phase of vaccine is complete.
27
The BRIDGE,Cell-Culture-Based Vaccine Production: Rino Rappuoli p 26-27,

28. KILLED / INACTIVATED VACCINE
• The term killed generally refers to bacterial vaccines, whereas
inactivated relates to viral vaccines (Levine et al., 1997).
• For viruses, the outer virion coat should be left intact but the
replicative function should be destroyed.
• Virus inactivation works by one of the following two
mechanisms:
- By attacking the viral envelope or capsid and destroying
its ability to infect or interact with cells.
- By disrupting the viral DNA or RNA and preventing
replication.
• Killed vaccines may take the route of heat or chemicals (Turner
et al., 1970).
28
The Biomedical Engineering Handbook: Second Edition. Ed. Joseph D. Bronzino ,Aunins, J. G., Lee. A. L.,Volkin, D. B. “Vaccine Production.”

29. Chemical treatment -
• The chemicals used for killed vaccines include formaldehyde
or beta-propiolactone (Lo Grippo, 1960; Gard, 1960).
• The traditional agent for inactivation of the virus is formalin
(Weil & Gall, 1940; Kim & Sharp, 1967).
• The agent is chosen for effectiveness without destruction of
antigenicity.
• For whole organisms, the inactivation abolishes infectivity.
• For antigens such as the diphtheria and tetanus toxins,
formaldehyde treatment removes the toxicity of the antigen
itself as well as killing the organism.
• Such detoxified antigen, called toxoids, are safe for use in
vaccines.
29
Recombination and Point Mutations in Type G Rotavirus Strains: The Challenges of Vaccine Development, Abid Nabil Ben Salem1, Rouis Zyed1, Buesa
Javier2 and Aouni Mahjoub1

30. Solvent/detergent (S/D) inactivation –
• Effective with lipid-coated viruses.
• The detergents used in this method, Disrupts the interactions
between molecules in the lipid coat , rendering the coat
dysfunctional and impeding replication.
• Most enveloped viruses cannot live without their lipid coating,
so they die when exposed to these detergents. Other viruses
may still live, but they are unable to reproduce, rendering them
non-infective.
• The detergent typically used is Triton-X 100.
30
The Biomedical Engineering Handbook: Second Edition. Ed. Joseph D. Bronzino ,Aunins, J. G., Lee. A. L.,Volkin, D. B. “Vaccine Production.”

31. By Heat -
A physical inactivation method that leaves intact virus, bacteria.
The heat inactivates the infectious agents, by destroying there
surface antigen.
Heat inactivation usually done at 56 C for 30 minutes.
e.g. A normal suspension of pertussis cells contains a thermolabile
toxin that is inactivated by heating at 56 C for 10 minutes.
Ultraviolet (UV) inactivation -
• UV rays can be used to inactivate viruses since virus particles
are small and the UV rays can reach the genetic material,
inducing the dimerisation of nucleic acids.
• Once the DNA dimerised, the virus particles cannot replicate
their genetic material. 31
The Biomedical Engineering Handbook: Second Edition. Ed. Joseph D. Bronzino ,Aunins, J. G., Lee. A. L.,Volkin, D. B. “Vaccine Production.”

32. LIVE WHOLE VACCINES:
Several methods have been used to attenuate viruses for vaccine
production.
a) Use of a related microorganism from another animal -
The earliest example was the use of cowpox to prevent smallpox.
b) Administration of pathogenic or partially attenuated
microorganism by an unnatural route -
The virulence of the virus is often reduced when administered by an
unnatural route.
32
Recombination and Point Mutations in Type G Rotavirus Strains: The Challenges of Vaccine Development, Abid Nabil Ben Salem1, Rouis Zyed1, Buesa
Javier2 and Aouni Mahjoub1

33. c) Passage of the microorganism in an "unnatural host" or
host cell
• The major vaccines used in man and animals have all been
derived this way.
• Example :
- the 17D strain of yellow fever was developed by passage in
mice and then in chick embryos
(Norrby, 2007).
- Polioviruses were passaged in monkey kidney cells
(Chezzi et al., 1998).
- Measles in chick embryo fibroblasts
(Katz, 1958).
33
Recombination and Point Mutations in Type G Rotavirus Strains: The Challenges of Vaccine Development, Abid Nabil Ben Salem1, Rouis Zyed1, Buesa
Javier2 and Aouni Mahjoub1

34. Formulation of vaccine
Finally, the vaccine is formulated by adding adjuvant, stabilizers, and preservatives as
needed.Other than microorganism or its part (antigen), a vaccine contain the following
components:
Component Purpose Example Example of Vaccines
Adjuvants Enhance the immune Aluminium salts Diphtheria-pertussis-tetanus
response to a vaccine (Alum) Diphtheria tetanus(DT)
DT combined with Hepatitis B
(HBV)
Haemophilus influenza B
Inactivated polio virus(IPV)
Hepatitis A (HAV)
Preservatives Prevent bacterial or Thimerosal Diphtheria-tetanus-acellular
fungal contamination of pertussis (DTaP)
vaccine Hepatitis B,
Haemophilus influenza type B
(Hib). 34

35. Compone Example of
Purpose Example
nt Vaccines
Stabilizers Protects vaccines from adverse Gelatine, 17D Yellow Fever
conditions such as freeze-drying Monosodium virus vaccine,
or heat, thereby maintaining a glutamate (MSG) Rabies,Varicella
vaccine’s potency
Inactivating agents - used to Formaldehyde Influenza virus,
inactivate bacterial products for Poliovirus, Diphtheria
toxoid vaccines, to kill and Tetanus toxins.
unwanted viruses and bacteria β-propiolactone
Rabies virus
that might contaminate the
Glutaraldehyde
Residuals vaccine during production Acellular pertussis
from Antibiotics - prevent bacterial DTaP-IPV/Hib
Neomycin,
manufactur contamination during
Streptomycin, Influenza, MMR
ing process manufacturing process
Polymyxin B
Influenza and yellow
Suspending fluids – Egg proteins
fever vaccines
kill or weaken the organism for
Hepatitis B vaccines
use in vaccines.
Yeast proteins

36. QUALITY CONTROL AND LOT RELEASE
Schedule of final product testing for a veterinary vaccine
Test Purpose of Test
Sterility Demonstrates that no live microorganisms are present in product
Safety Demonstrates that overdose of the product causes no harm
Residual toxicity Demonstrates that the product contains no material that can cause harm
Efficacy Demonstrates that each antigen in the product meets the recommended
guideline level in internationally recognised tests.
Increase in With live vaccines, there is concern that the organism might be shed from
virulence tests the host and transmitted to contact animals, causing disease if it retains
residual virulence or reverts to virulence.
All live vaccines should be tested for virulence by means of passage
studies.
Interference tests For products with two or more antigenic components, tests must confirm
that there is no interference between individual components, that is, one
PRINCIPLES OF
VETERINARY VACCINE component causing a decrease in the protective immunological response
PRODUCTION, OIE Terrestrial 36
Manual 2008
to another component.

37. Lot release
BATCH/SERIAL RELEASE FOR DISTRIBUTION :
Prior to release, the manufacturer must test each
batch/serial for purity, safety, and potency.
1. Batch purity test –
Purity is determined by testing for a
variety of contaminants.
Tests to detect contaminants are
performed on: master seeds, primary
cells, MCSs(Master cell stock), and
each batch of final product prior to
release. 37
PRINCIPLES OF VETERINARY VACCINE PRODUCTION, OIE Terrestrial Manual 2008

38. 2. Batch safety test -
Batches are considered satisfactory if local and
systemic reactions to vaccination with the batch to
be released are in line with those described in the
registration dossier and product literature.
3. Batch potency test -
Batch/serial potency tests, required for
each batch prior to release, are
designed to correlate with the host
animal vaccination–challenge efficacy
studies. 38
PRINCIPLES OF VETERINARY VACCINE PRODUCTION, OIE Terrestrial Manual 2008

39. Sampling :
Samples should be selected from each batch/serial
of product.
The selector should pick representative sample.
Filling , Packaging and Labelling
Once all procedures of vaccine production are completed, the
vaccine is blended, filled the doses into vials and packaged.
Which are then sealed and carefully inspected before labels are
applied to show the vaccine batch,lot numbers, and expiration
date.
Standards for labelling products will vary from country to
country.
Field tests (safety and efficacy)
Performance monitoring 39
PRINCIPLES OF VETERINARY VACCINE PRODUCTION, OIE Terrestrial Manual 2008

40. Processes of Egg-Based Vaccine Production
Embryonated
chicken eggs Inoculation Incubation
The upstream A seed ampoule This is followed by a 3
process, begins is used to day incubation period
with embryonated inoculate the during which the virus
eggs brought in chick eggs grows to ensure that
on a daily basis during the sufficient quantities
from biosecure inoculation can support further
flocks. phase. manufacturing.
Harvest (pool Candling
allantoic fluid)
After 3 days, all of the eggs are
The allantoic fluid is candled to make sure there are no
then harvested; a low- cracks or contamination; the eggs
Clarification speed clarification are then chilled to 2° to 8°C to
process follows. constrict vessels and make
harvesting easier. 40
The BRIDGE, Egg-Based Production ofInfluenza Vaccine:James T. Matthews page 21

41. 1st inactivation Filtration Concentration
The first step in the
Zonal centrifugation
downstream Extensive filtration and concentration
process is steps yield a concentrate, which is
inactivation, which then loaded onto zonal centrifugation
involves the equipment.
addition of formalin
to inactivate the
virus.
Triton fragmentation
2nd inactivation Centrifugation
Ultrafiltration The material is then The first purified
clarified by bulk virus, which is
An ultrafiltration centrifugation to recovered from the
(UF) step is remove large centrifugation
followed by terminal particulates and process, is split in
sterile filtration to Sterile filtration treated with a fragmentation
generate one formalin in a step by treatment
monovalent bulk second inactivation with Triton
concentrate. step. detergent. 41
Final product The BRIDGE, Egg-Based Production of Influenza Vaccine:James T.

42. Cell Culture Based Vaccine Production
centrifugation
virus
cell (production
seed)
   filtering
Cell culture Inoculation Harvest Bulk Purification

Add
Adjuvant Stabilizer
   
Preservative
Packaging Labeling Inspection Filling Formulation
42
ccines / [edited by] Stanley A. Plotkin, Edward A. Mortimer, Jr. Influenza Vaccines (Tina Kröber, Thomas Jarosch, and Laura Fischer) Max-Planck-Gesellschaft, Münche

43. Thank
you
43

44. 44

45. • Production of Freeze-dried Brucella abortus Strain 19 Vaccine
using Cells produced by Continuous Culture† K. J. Boyce, A. W.
Edgar ,Journal of Applied Microbiology, Volume 29, Issue 2, pages
401–408, August 1966
• The tularaemia vaccine: review. Sandstrom, G.
Journal of Chemical Technology & Biotechnology. Vol. 59, no.
4, pp. 315-20. Apr. 1994
The live vaccine can be produced in fermenter cultures and the
bacteria have to undergo quality controls for each batch of vaccine.
It is important to estimate the amount of non-immunogenic bacteria
in each batch and to disclose the immunogenicity of the vaccine in
man.
45