vaccine production,techniques

1. RABIES
VACCINE
-B.HARISHA
4thyear(CLBM)

2. INTRODUCTION
 It is primarily a ZOONOTIC disease of warm blooded.
 Acute rapid progressive &highly fatal viral disease of CNS
 Transmitted to man by the bite of rabid animals.
 Non-bite exposure:aerosol generated in lab.
corneal transplantation.
 Human to human transmission is rare.
CAUSATIVE AGENT: Lyssa virus type 1belongs to the family RHABDOVIRUSES
which is a bullet shaped neurotropic single stranded RNA Virus.

3. ANTIRABIES VACCINES
 In 1883,Louis Pasteur
performed the first successful
Human Antirabies virus(HAP)
 The original vaccine was the
suspension of nervous tissue of
animals infected with fixed
rabies virus.

4. STEPS IN VACCINE PRODUCTION

5.  SELECTION:It involves the viruses that
is derived from seed lot system.
 GROWING:Methods used are 1.cell
culture 2. Bird embroyos 3.Live animal
inoculation 4.Transgenic animals.
 ISOLATION &PURIFICATION:It
involves 1.centrifugation 2.Filteration
3.chromatography

6. SEED LOT SYSTEM
A seed lot
system is a system in
which successive
products of the batch
are derived from the
same master seed lot
at a given passage
level.For routine
production,a working
seed lot is prepared

7.  INACTIVATION: (virus INACTIVATION)It can be done by dismantling
a virus’s ability to infect cells without actually eliminating the virus.
 METHODS: 1.Solvent/detergent inactivation. Eg:TRITON-X 100.
2.Pasteurization(60 C for 10 hrs)
3.Acidic pH inactivation (pH 4 incubation btw 6 hrs and 21
days)
4.UV inactivation(diminish replication)
5.Inactivation by extraction (nucleic acid….)

8. FORMULATION: it includes
1.suspending fluids(sterile water,saline or fluids
containing protein)
2.preservative(thimerasal) and
stabilizer(albumin,phenol,glycine.MSG-stimulator,
antibiotics)
3.inactivating agent(beta propiolactone)
4.adjuvants or enhancer

9. TECHNIQUES
By utilizing free living animals
By utilizing fertilized egg
Tissue culture method

10. BY UTILIZING FREE LIVING
ANIMALS
STAGE 1- selection of animals
STAGE 2-Inoculation(intracerebral)
STAGE 3-Incubation(24hrs)
STAGE 4-Harvesting (homogenized with Nacl)
STAGE 5-Purification and inactivation

11. A) Neural vaccines:
 FERMI VACCINE:fixed virus is grown in sheep brain& treated with phenol
at 22 C.some residual virus is present.
 SAMPLE VACCINE:Fixed virus is grown in sheep brain and inactivated
with phenol at 37 C leaving no residual virus.widely used.
 BETA PROPIOLACTONE VACCINE:The sheep brain vaccine is
inactivated by this which is more antigenic.
 INACTIVATED VACCINES:Prepared in brain of suckling
animals(rat,mice,rabbit).risk of neurological complications.

12. BY UTILIZING FERTILIZED
EGG
 DUCK EGG VACCINE:In this fixed virus is adapted for
growth in duck embryos and inactivated with
Betapropiolactone.
Not effective as brain vaccines for post
exposure treatment.
Occasional neurological complications.

13.  LIVE ATTENUATED CHICK EMBRYO
VACCINE:In this flury strain of attenuated
rabies virus is used.Two types available they
are 1.Low egg passage(40-50 egg passage)
2.High egg passage (180th passage for
cattles..used for human also-preimmunization)

15. TISSUE CULTURE METHOD
 It is a great advance in prophylaxis.
 More potent and cheap and much safer than the other.
 There are two types of tissue culture vaccines they are
1.Human diploid cell vaccine(HDCV)
2.Non human source vaccine

16.  HUMAN DIPLOID CELL VACCINE: It is prepared by propogating
the rabies virus in human diploid fibroblast cells.
Safe and high potent but not in use for pre and post
immunization.
 NON HUMAN SOURCE: The viruses grown in hamster kidney
cells,chick embryo fibroblast,dog kidney cells,vero cells(kidney
epithelial cells of African green monkey- used in cell culture)
Hypersensitivity: headache, palpitation,giddiness,
diarrhoea,itching,pain,redness and swelling at site of reaction,
anaphylactic reaction.

18. EVALUATION OR TESTS OF
VACCINES
 Increase in virulence tests
 Assessing risk to the environment.
 Interference test
 Consistency of production
 Stability tests
 Batch/serial purity test
 Batch/serial safety test
 Batch/serial potency test
 Other tests(moisture,pH,residual inactivate, physical stability etc…)

19. SAMPLING: Samples should be selected
from each batch /serial of product.
LABELLING:Standards for labelling
products will vary from country to country

22. Thank
you!