Downstream processing of vaccine

(e.g. Influenza & Tetanus)

Modern College Of Arts , Science & Commerce.Shivajinagar, Pune-05.

Presented By-NIYAMAT M. PANJESHA

SNEHA SAWLEYUGENDRA PATIL

ASEMINAR ON

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.It was in 1796 Jenner experimentally tested

this observation.The cowpox infection

could protect these people from infection of smallpox.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.

HIST

ORY

TypesTraditional

Live attenuated Organism

Killed

Toxoid

RecombinantSub-unit

Conjugate

Vector

ExperimentalDNA vaccine

Edible vaccine

Examples of VaccinesSr. No.

Vaccine Organism Disease

1 Live Attenuated Bacterial Vaccine

•Bacillus anthracis•Salmonella typhii

•Aanthrax•Typhoid

2 Inactivated Bacterial VaccinesCapsular AgCell wall AgToxoid Ag

•Neissseria meningitides•Vibrio Cholera•Clostridium tetani

•Meningitis•Cholera•Tetanus

3 Recombinant VaccinesBacterialProtozoalViral

•Clostridium tetani•Plasmodium vivax-H1N1

•Tetanus•Malaria•Influenza

4 DNA vaccines •Malaria

Steps In Vaccine Production

SELECTING THE STRAIN FOR VACCINE PRODUCTION

GROWING THE MICRO-ORGANISM

ISOLATION & PURIFICATION OF MICROORGANISM

INACTIVATION OF ORGANISM

FORMULATION OF VACCINE

QUALITY CONTROL AND LOT RELEASE

Up

stream

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ream

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Two modes: Seasonal Epidemic and Pandemic Influenza

Types of Vaccine Production:1.Egg based(Traditional)2.Cell line based

Three forms of vaccine:A.WholeB.SplitC.Subunit/Surface Antigen

Influenza Vaccine

Live Virus Vaccine Production using

MDCK cell line on Cytodex3 micro carrierUpstream 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.

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.

DOWNSTREAM PROCESSING

TETANUS VACCINE

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.

Production strain

Revival of the production strain

Preparation of seed culture

Production of tetanus toxin by Static

Culture Method

Harvesting of Toxin

Determination of antigenic content

Detoxification of the Toxin

Partial purification of tetanus toxin

Purification of crude tetanus

toxoid

Tetanus Vaccine Production

12

Alum

Production strain Seed culture Fermentation Purification

Adjuvant

FillingLabeling Packaging

Inoculation

Inspection

Ultrafilteration

Glycine soln Formaldehyde soln

Monosodium glutamate

Inactivation

Thimersol

Stabilizer

Preservative

Dilutant

REFERENCESNielsen, 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 VaccinesPropagation and purification of influenza virus.

THANK YOU

QUESTIONS, CRITICISMS, AND SUGGESTIONS?

snehasawle@rediffmail.com niyamatpanjesha@yahoo.com