Production and purification of rabbis vaccine

2.  Roll: 461
 Batch : 16th
Department of pharmacy
World university of Bangladesh

3.  Rabies is a zoonotic disease which is caused by a virus.
 Rabies virus (RABV) belongs to the type species of the genus Lyssavirus within the family
Rhabdoviridae
 Rabies infects domestic and wild animals, and is spread to people through close contact with
infected saliva through bite, scratch, aerosols etc

4.  Dogs,
 Bats,
 wild cats,
 Jackals,
 wolves etc

10.  RABV infection starts at a peripheral site
 The virus enters unmyelinated nerve terminals and migrates by retrograde axonal transport to
the neuronal cell body.
 After replication in the cell body of the primary neuron, infection proceeds via retrograde
axonal transport and trans-synaptic spread through several neurons and reach the CNS
 Then it infects the acinar cells which releases the virus into the oral cavity by passing on to the
salivary glands.

12.  The incubation period varies from 2 weeks to 6 years depending on the amount
of virus in the inoculum and the site of inoculation
 First clinical symptom is usually neuropathic pain at the wound site.
 Encephalitis
 Virus grows to high titers in the salivary glands
 Negri bodies appear in neuron cell bodies
 Clinical spectrum
 Prodrome - nausea, headaches, fever, sore throat, photophobia
 Acute neurologic phase - nervousness, hallucinations, behavioral anomalies,
salivation, perspiration, hydrophobia, Coma and death.

13. Vaccines produced by employing adult animal nerve tissue
Presence of residual live virus: vaccines such as the
Presence of residual live virus: vaccines such as theFermi vaccine
which contain live virus because of deficient inactivation with
phenol, are considered mixed vaccines should not be used
Post-vaccinal encephalomyelitic reactions: certain vaccines from
this group including the Semple andHempt vaccine types are fully
inactivated but, despite fulfilling potency requirements, severe post-
vac-cinal encephalomyelitic reactions have occurred during

14. Low antigen content per dose of vaccine. This feature made long
treatments necessary with a great number of inoculations. These
vaccines, generally sold in a liquid vehicle, had low stability of
approximately 6 months

15. An adaptation of rabies virus strains in duck embryos. enabled the
development of much safer rabies vaccines for human use,
diminishing neuro paralytic post vaccinal encephalomyelitic
reaction
The incidence of neurological reactions is much lower in human
vaccines produced in duck embryos than in those produced in nerve
tissue. However, local reactions are quite common, reaching 70% in
prophylactic vaccination and 100% in complete,14-dose rabies
treatments . Immunogenicity of this vaccine type is a matter of
debate due to the low antibody levelpro vided in some cases

16.  Flury and Kelev fixed virus strains were adapted to chick embryo
through repeated passaging yielding attenuated viral strains .
Vaccines produced with attenuated virus in chicken embryos are
used in dogs, cats and cows
Very high quality embryonated eggs must be free from all kinds of
adventitious virus and bacteria for use in vaccine production

17. The encephalitogenic factor likely to be responsible for post
vaccinal neurological accidents is present in adult animal nerve
tissue, but is negligible or absent in certain suckling animals.
Together with the higher viral productivity achieved in new-born
animal nerve tissue, this finding has
fostered development of vaccines employing suckling animals .
The first vaccine of this type was developed by Fuenzalida and
Palacios in suckling mouse brain, employing three strains of fixed
rabies virus

18. The first group comprises vaccines manufactured in primary
mammalian cell cultures, such as hamster kidney dog kidney and
fetal calf kidney or in avian cell cultures such as chicken embryo
and quail embryo following Kiss ling’s methodology
Second group:
Second group vaccines are produced in diploid cells of regular
cariogenicity and duplication , mainly of human origin Some
vaccines of animal origin likewise pertain to this group

19. The third group includes vaccines developed in hetero ploid cell
culture as the Vero line
The development of rabies vaccines for veterinary use, which are
usually employed for prevention , was more widely favored than
that of vaccines for human use.
This preference is due not only to a greater demand for the former
but also because production requirements are less stringent for
veterinary vaccines. features of production processes for these
rabies vaccine

20. Used the AKTA purifier system and different chromatography
columns which are Capto Core 700 column and the CIMmultus
QA-8, an anion exchange advanced composite column.
300 mL of clarified and inactivated viral harvests were concentrated
6 fold by tangential flow ultrafiltration on Pellicon cassette Biomax
XL with a cut-off 100 kDa, then purified on Capto Core 700
column as described in Materials & Methods.

21.  After purification, the flow through and 20 fractions of 1 mL were analyzed by SDS-PAGE and
ELISA to determine glycoprotein content.

22. The results are indicated in absorbance at 280 nm showed a major
peak at fraction F3 related to the impurities coming from the loaded
sample.
The collected fractions were also analyzed by SDS-PAGE; intense
bands were seen in F3 fraction
this correlates with absorbance at 280 nm level