process involved in production of tetanus and influenza vaccine.

1. A
SEMINAR ON
Downstream processing
of vaccine
(e.g. Influenza & Tetanus)
Presented By-
NIYAMAT M. PANJESHA
SNEHA SAWLE
YUGENDRA PATIL
Modern College Of Arts , Science & Commerce.
Shivajinagar, Pune-05.

2. What are Vaccines?
• A vaccine is a biological preparation that improves
immunity to a particular disease.
• Name derived from the word vacca which means cow.
• Can be- 1) Prophylactic. 2) Therapeutic.
Edward Jenner observed that the farmers and milkmaids working with cows
developed a mild form of small pox called cowpox or vaccinia.
The cowpox infection could protect these people from infection of smallpox.
It was in 1796 Jenner experimentally tested this observation.
He inoculated an 8 yr old boy with exudates from a cowpox
lesion and he repeated it twice with a gap of some weaks.
Jenner noticed that smallpox did not develop in this boy.
This was 1st discovery of principle of vaccination.

3. Live attenuated
Organism
Killed
Traditional
Toxoid
Sub-unit
Types Recombinant Conjugate
Vector
DNA
Experimental vaccine
Edible
vaccine

4. Examples of Vaccines
Sr. Vaccine Organism Disease
No.
1 Live Attenuated Bacterial •Bacillus anthracis •Aanthrax
Vaccine •Salmonella typhii •Typhoid
2 Inactivated Bacterial
Vaccines
Capsular Ag •Neissseria meningitides •Meningitis
Cell wall Ag •Vibrio Cholera •Cholera
Toxoid Ag •Clostridium tetani •Tetanus
3 Recombinant Vaccines
Bacterial •Clostridium tetani •Tetanus
Protozoal •Plasmodium vivax •Malaria
Viral -H1N1 •Influenza
4 DNA vaccines •Malaria

5. Steps In Vaccine
Production

6. processing
SELECTING THE STRAIN FOR
Upstream
VACCINE PRODUCTION
GROWING THE MICRO-ORGANISM
ISOLATION & PURIFICATION OF
MICROORGANISM
Downstream
processing
INACTIVATION OF ORGANISM
FORMULATION OF VACCINE
QUALITY CONTROL AND LOT RELEASE

7. Influenza Vaccine
Two modes: Seasonal Epidemic and
Pandemic Influenza
Types of Vaccine
Production:
1.Egg based(Traditional)
2.Cell line based
Three forms of vaccine:
A.Whole
B.Split
C.Subunit/Surface Antigen

8. Live Virus Vaccine
Production using
MDCK cell line on
Cytodex3 micro carrier
Upstream Process:
0.5X106 cells/ml(at 100r.p.m.
for 48hrs.)
At 2-2.5X106 cells/ml infect
with Viral inoculum and carry
out for 24hr.

9. DOWNSTREAM
PROCESSING
1.Clarification:
ULTA Prime GF capsule
Filtration(0.6µm)
2.Diafiltration(0.4µm) in
buffer[20mM Tris+0.5M NaCl,
pH7.5]
3.DNA binding Anion Exchange
Chromatography: To remove gDNA
4.Sterile Filtration(0.2µm):
Stainless steel filter precoated with
buffer.

10. Tetanus is an acute and highly fatal disease caused by
exotoxins produced by the bacterium Clostridium.tetani.
Cl. tetani produces two exotoxins- tetanolysin and
tetanospasmin.
Tetanus toxoid is one of the most immunogenic
antigens available for the protection against an infectious
disease in the world.
Tetanus Vaccine produced by anaerobic fermentation.

11. Tetanus Vaccine Production
Revival of the Preparation of seed
Production strain
production strain culture
Production of
tetanus toxin by Determination of
Harvesting of Toxin
Static Culture antigenic content
Method
Detoxification of the Partial purification Purification of
Toxin of tetanus toxin crude tetanus toxoid

12.    
Production strain Inoculation Seed culture Fermentation Purification

Monosodium   Formaldehyde  Glycine 
glutamate Alum soln soln
Stabilizer Adjuvant Dilutant
Inactivation Ultrafilteration

Thimersol
   
Preservative Inspection Labeling Packaging
Filling
12

14. REFERENCES
Nielsen, P. A.1967. Large-Scale Production of Vaccine. Applied Microbiology.
Ozutsumi K, Sugimoto N, and Matsuda M. 1985. Rapid, Simplified Method for
Production and Purification of tetanus toxin. Applied and environmental
microbiology.
 Chandani Payal and Tejpal Kashyap, 2011. Production, Optimization of
Detoxification and Ammonium Sulphate Precipitation of Ultrafiltered Tetanus
Toxin. Recent Research in Science and Technology, 3(11): 49-54.
 Nduka Okafor, Modern industrial microbiology and biotechnology.
U.Satyanarayan, A Text book of Biotechnology.
Michael J.Waites, Industrial Microbiology: An Introduction.
GE Healthcare Life Sciences, Application note 28-9843-41 AA Vaccines
Propagation and purification of influenza virus.

15. QUESTIONS, CRITICISMS, AND
SUGGESTIONS?
THANK YOU
snehasawle@rediffmail.com
niyamatpanjesha@yahoo.com