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Viral Vaccine
Production
SOURABH SANTOSH GURAV
MTECH BIOPROCESS TECHNOLOGY
INSTITUTE OF CHEMICAL TECHNOLOGY
IMPORTANT AREAS FOR A VIRAL
VACCINE
• Five important areas for a viral vaccine are -
(1) the choice of the cell substrate
(2) the choice of the virus construct
(3) bioreactor technology
(4) purification
(5) final product stabilization.
Cell Substrates
• Human diploid cell lines: Medical Research Council-5 (MRC-
5) & Wistar Institute-38 (WI-38)
• Continuous cell lines : Vero and Madin Darby Canine Kidney
(MDCK)
• recombinant “designer” cell lines: PER.C6 and CAP
• plant-based recombinant protein production (Not
commercial)
VACCINE TYPE REMARK EXAMPLES
• Inactivated Vaccine or
killed vaccine
Pathogens that have been grown in culture and
then lose disease producing capacity
Hepatitis A, whole cell
Pertussis
• live vaccines Pathogens are still alive but are almost attenuated,
that is, weakened
Oral Polio Vaccine,
Rotavirus, Tuberculosis
(BCG) etc
• Subunit vaccine Contains a fragment of the pathogen and elicits an
appropriate immune response
• Diphteria, acellular
Pertussis, Subunit influenza
• Virus - like - particle
Vaccine
Molecules that mimic viruses but are not
infectious
Human Papilloma Virus
(HPV)
• Viral Vector Vaccine Uses a chemically weakened virus to transport
pieces of the pathogen
• DNA , RNA Vaccine Uses a man-made copy of a natural chemical
called messenger RNA (m RNA) or DNA to produce
an immune response
Bioreactor Technology
• Attachment-dependent modalities such as t-flasks, cell factories,
and roller bottles
• At industrial scale –
i. Small spheres (microcarriers) kept in suspension
ii. 6000L scale
• Single-use bioreactors
i. Facilitates multiproduct manufacturing in facilities and reduces
change-over times between batches
Microcarrier Cell System
Hollow fibre Membrane
1.CELL
EXPANSION
2.VIRUS
PROPOGATION
(BIOREACTOR)
3.Removal of cell
debris and large
particles
4.CONCENTRATION
5.Separation of virus
from small molecular
compounds
6.Virus
inactivation
formaldehyde
5.Formulation
Sterile filtration,
mixing with other
strains
typical viral vaccine production process
• Harvesting of the virus by centrifugation
and/or filtration
• Purification via chromatograph
• Inactivation as needed with chemical
additives
• Final conditioning with tangential flow
filtration
• Size-exclusion chromatography :purification
of whole virus
• Flow-through chromatography :impurities in
the feed are adsorbed to the resin
Cell Culture Roller Flask System
Virus Stabilization
• addition of complex stabilizers and via lyophilization for live
viral vaccines
• enable distribution
• need to supply a separate diluent in addition to the dry
powder
• spray drying have been investigated
• spray-drying
mechanism
STORAGE
live-virus vaccines :tolerate freezing temperatures and rapidly
deteriorate once defrosted
inactivated vaccines :require stable temperatures of 2-8⁰C
Viral vaccine production (5 MIN READ)

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Viral vaccine production (5 MIN READ)

  • 1. Viral Vaccine Production SOURABH SANTOSH GURAV MTECH BIOPROCESS TECHNOLOGY INSTITUTE OF CHEMICAL TECHNOLOGY
  • 2. IMPORTANT AREAS FOR A VIRAL VACCINE • Five important areas for a viral vaccine are - (1) the choice of the cell substrate (2) the choice of the virus construct (3) bioreactor technology (4) purification (5) final product stabilization.
  • 3. Cell Substrates • Human diploid cell lines: Medical Research Council-5 (MRC- 5) & Wistar Institute-38 (WI-38) • Continuous cell lines : Vero and Madin Darby Canine Kidney (MDCK) • recombinant “designer” cell lines: PER.C6 and CAP • plant-based recombinant protein production (Not commercial)
  • 4. VACCINE TYPE REMARK EXAMPLES • Inactivated Vaccine or killed vaccine Pathogens that have been grown in culture and then lose disease producing capacity Hepatitis A, whole cell Pertussis • live vaccines Pathogens are still alive but are almost attenuated, that is, weakened Oral Polio Vaccine, Rotavirus, Tuberculosis (BCG) etc • Subunit vaccine Contains a fragment of the pathogen and elicits an appropriate immune response • Diphteria, acellular Pertussis, Subunit influenza • Virus - like - particle Vaccine Molecules that mimic viruses but are not infectious Human Papilloma Virus (HPV) • Viral Vector Vaccine Uses a chemically weakened virus to transport pieces of the pathogen • DNA , RNA Vaccine Uses a man-made copy of a natural chemical called messenger RNA (m RNA) or DNA to produce an immune response
  • 5. Bioreactor Technology • Attachment-dependent modalities such as t-flasks, cell factories, and roller bottles • At industrial scale – i. Small spheres (microcarriers) kept in suspension ii. 6000L scale • Single-use bioreactors i. Facilitates multiproduct manufacturing in facilities and reduces change-over times between batches
  • 7. 1.CELL EXPANSION 2.VIRUS PROPOGATION (BIOREACTOR) 3.Removal of cell debris and large particles 4.CONCENTRATION 5.Separation of virus from small molecular compounds 6.Virus inactivation formaldehyde 5.Formulation Sterile filtration, mixing with other strains typical viral vaccine production process
  • 8. • Harvesting of the virus by centrifugation and/or filtration • Purification via chromatograph • Inactivation as needed with chemical additives • Final conditioning with tangential flow filtration • Size-exclusion chromatography :purification of whole virus • Flow-through chromatography :impurities in the feed are adsorbed to the resin Cell Culture Roller Flask System
  • 9. Virus Stabilization • addition of complex stabilizers and via lyophilization for live viral vaccines • enable distribution • need to supply a separate diluent in addition to the dry powder • spray drying have been investigated
  • 11. STORAGE live-virus vaccines :tolerate freezing temperatures and rapidly deteriorate once defrosted inactivated vaccines :require stable temperatures of 2-8⁰C