Bioprocess method pf vaccine production.

1. Industrial Manufacturing Process for Vaccines
Development
- Beh Yoke Leng (0338291)
- Ngan Boon Keat (0333894)
- Ng Xian Gui (0337937)
- Chu Sin Ton (0338668)

2. Introduction:
Vaccines:
- Is a preparation of killed microorganisms, living attenuated
organisms, or living fully virulent organisms that is
administered to produce or artificially increase immunity to
a particular disease
Type of Vaccine
Inactivated
Attenuated
Toxoid
Recombinant Subunit
Conjugate Polysaccharide-Protein

3. Stages of vaccine production
Upstream Process
1. Selecting the strain for vaccine
production
2. Growing the microorganisms
(Propagation)
● Manufacturing begin with a small amounts of
specific virus or bacteria (shall be derived from a
Seed Lot System)
● The seed (strain) must be kept under “ideal”
conditions,usually frozen to pprevent the virus
from becoming either stronger or weaker.
● The choice of the seed(strain) is depends on a
number of factors including the efficacy of the
resulting vaccine and its secondary effect
Growing bacteria
(explanation in method part)
Growing viruses
(explanation in method part)

4. Downstream
Process
● Virus inactivation
● Ultraviolet inactivation
● Inactivation by nuclei acid /membrane/capsule
extraction
4. Inactivation of organisms
3. Isolation and purification of microorganisms

5. 5. Formulation of vaccine
6. Quality control and lot release
Prior to release,the manufacturer must test
each batch/serial for purity,safety and potency.

6. Method of growing microorganism
Growing bacteria
Continuous cultureBatch culture
● Close system
During fermentation
○ no fresh nutrient &
microbes pump in
○ nothing pump out
● Culture decline due to the
limiting factor; eg, nutrient
● Product can be harvested
from the culture only after
fermentation
● Open system
During fermentation
○ continual provision of
nutrient
○ continual removal of
waste
● Culture can be grown
indefinitely by keeping the
condition in optimal value
● Product can be harvested
from the culture
continuously.
Chemostat

7. Growing viruses
Cell culture Embryonated egg
Live animal
inoculation
Trangenic animal
● Primary cell culure
○ directly from animal/
human tissue
○ only can subculture
once/ twice
● Secondary cell culture
○ derived from human
fetal tissue
○ can subculture 20-50
times
● Continuous cell line
○ able to propagate
indefinitely
○ derived from tumor of
mutated cell line
● A number of
viruses
propagated in
embryonated
chicken egg
● A hole is drilled in
the cell and virus
was injected in
appropriate site
for its replication.
eg, yolk sac →
Herpes simplex
virus
● Experiment animal
are obligatory for
studying virus
pathogenesis
● Mice are most
widely used
experimenal
animal
● Insertion of the DNA
of the virus genome
● Virus RNA and
protein expressed in
somatic cells
● Modification allow
target the gene to
specific cell
● Best example is the
creation of
transgenic mice
expressing hepatitis
B virus

8. Example of vaccine production
Hepatitis B vaccine - Recombinant yeast method
Strain selection
Preparation of vectors carrying DNA
sequence for HBsAg and
antigen expression in the yeast
Saccharomyces cerevisiae or H.
polymorpha
Cell culture/fermentation
● 5-L seed fermenter used to inoculate the
50-L main fermenter
● Carried out in synthetic medium feeding of
glycerol and methanol in two different
phases.
● Recombinant hepatitis B surface antigen
(HBsAg) produced in yeast cells grown in a
complex medium of extract of yeast, soy
peptone, dextrose, amino acids, and
mineral salts

9. Isolation of microorganisms
Purification
Released from yeast cell by cell distruption
● Ultrafiltration and gel permeation chromatography
● Formalin inactivation
Formulation of vaccine
● Absorbed into aluminium hydroxide
● None preservative method
Cell distruption
Cell distrupted by ion exchange, ultra filtration, and
gel filtration steps

10. Potential Challenges: Future Development:
Vaccine Hesitancy:
- Reluctance/refusal of vaccine as the result of
complacency, inconvenience, or doubt about
their effectiveness (Alex and er, 2020).
Ethical Considerations:
- Human clinical trials for testing new vaccine
(WHO,2013)
Inadequate Preclinical Data:
- lack of detailed information protective
correlates of immunity contribute to product
failure in clinical trials.
New Molecular Manipulation Development:
- New methods of molecular manipulation
enables faster development.
Refinements of existing vaccines:
- Reduces side effects that prevents mass
immunization.

11. References:
- ENGLISH ONLY EXPERT COMMITTEE ON BIOLOGICAL STANDARDIZATION Human Challenge Trials for
Vaccine Development: regulatory considerations. (2016). [online] Available at:
https://www.who.int/biologicals/expert_committee/Human_challenge_Trials_IK_final.pdf [Accessed 8 Jul. 2020].
- Alex, S. and er (2020). 5 vaccination challenges you need to know. [online] ONE. Available at:
https://www.one.org/international/blog/5-vaccination-challenges/ [Accessed 8 Jul. 2020].
- Oyston, P. and Robinson, K. (2012). The current challenges for vaccine development. Journal of Medical
Microbiology, 61(7), pp.889–894.
- W J McAleer, E B Buynak (2012). Human Hepatitis B Vaccine From Recombinant Yeast. Available at:
https://pubmed.ncbi.nlm.nih.gov/6318124/
- Gotthard Kunze, Georg Melmer, Gerd Gellissen (2008). Recombinant Vaccine Production in Yeast. Available at:
https://www.biopharminternational.com/recombinant-vaccine-production-yeast?id=&sk=&date=&pageID=3
- Vaccine production techniques 2020 Slidesharenet. viewed 9 July 2020, <https://www.slideshare.net/Drvijayata/vaccine-production-
techniques>.