This document discusses quality assurance of vaccines. It covers in-process quality testing such as staining, inactivation, sterility, and safety testing. Finished product testing includes potency testing through in vivo and in vitro methods. Vaccine stability depends on factors like purity, formulation, storage conditions and is assured through real-time, accelerated, and stress testing. Proper cold chain supply from manufacturer to vaccination site is crucial to maintain vaccine quality. Guidelines are provided for storage of heat, freeze and light sensitive vaccines. Shake testing can detect freezing damage in vaccines.

1. Quality assurance of Vaccines
Prepared by Maazullah
Gmail: pharmacistfuture7@gmail.com
Department of Pharmacy Shaheed Benazir Bhutto University Dir Upper
KP Pakistan
Subject : Pharmaceutical Quality Management
Subject Incharge :Dr Najm ur Rehman
References
• http://www.alliedacademies.org/microbiology-current-research/
• http://www.cdc.gov/vaccines/conversations
• https://doi.org/10.1080/01652176.2000.9695063
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2. Vaccines Quality Assurance
Contents
:
Vaccine definition and types :Page 3-4
In process quality testing :Page 4-11
Finished product testing :Page 12-15
Vaccine stability :Page 16-17
Cold chain supply :Page 18
Heat and freeze sensitive vaccines :Page 19-21
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3. Vaccine and types of vaccine
• Vaccines: Vaccine is a biological preparation that consists of either a
whole organism or a part of it against which immunization has to be
achieved.
Types of vaccine
1)Live attenuated vaccine (no pathogenicity but having immunogenicity
the ability of organism to provoke an immune response )like
measles,mumps,rubella ,smallpox
2)Inactivated /killed vaccine :made from micro-
organisms(viruses,bacteria ,other)have been killed by physical or
chemical processes .like inactivated polio vaccine and Rabies vaccine
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4. • Toxoid vaccine : Use of toxin a toxic substance produce by various
bacteria that cause a disease. In this case toxin has made harmless
but can induce an immune response when administered to the body .
like Tetanus toxoids and Diphtheria toxoids
• Subunit vaccine : It contain fragments /part of pathogen such as
proteins or polysaccharides or surface spikes instead of whole
organism.like Hepatities B vaccine
Quality assurance of vaccines : Part of quality management focused
on providing confidence that quality requirements will be fulfilled and
assured that the quality product will be made.
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5. • In process quality testing
Definition
• In process quality control is a process of
monitoring critical variables of
manufacturing process to ensure a quality
of the final product.
• In-process manufacturing controls are
established and documented by quality
control and production personnel to
ensure that quality of the product is
within the acceptable standard range.
• Staining test
• Inactivation test
• Sterility test
• Freedom from abnormal
toxicity/Generally safety test
Mainly to provide assurances of both the probable efficacy and safety of every batch of every product.
There are two ways
1. In-process control
2. Final product control
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6. • 1. STAINING TEST
Technique was developed by Dr Christian Gram in 1884.
Approximately 10 mL of the test sample is centrifuged in a pointed centrifuge tube
at approximately 2,000 rpm for 30 minutes. The sediment or the bottom portion
is spread on a slide glass, dried and heat-fixed over a flame. The smear is then
stained by the Gram’s Method and, unless otherwise specified, examined
microscopically. Criteria for judgment. No bacteria shall be observed other than
those defined in the individual monographs.
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7. 7

8. Freedom from abnormal
toxicity/General safety test
The test was developed in early 1900s to detect possible toxic
contamination derived from the manufacturing processes of the product.
Test in mice
Take 5 healthy mice weighing 17-22g. Inject one human dose 1 ml Intra-
peritoneally .Observe the mice for 7 days If more than one animal dies
preparation fails the test If one animal dies repeat the test. Preparation
passes the test if no animal dies in the second group.
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9. Test in guinea pigs
• Take 2 healthy guinea pigs (250-350 g).
• Inject one human dose 5 ml Intra-peritoneally.
• Observe guinea pigs for 7 days If more than one animal dies/shows ill
health preparation fails the test If one animal dies/ shows ill health
repeat the test Preparation passes the test if no animal dies/ shows ill
health in the second group .
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10. Sterility test
1. Immersion (Direct Inoculation)
• For bacteria – Fluid thioglycolate media use for both aerobics
anaerobic bacteria
Media preparation – sample transferring into media –Incubation 30-
35 c according to IP for 7 days
• For fungi - proceed as described in the test for bacteria but use
soyabean casein digest medium and incubate the plates at 20 °C to
25 °C for 14 days
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11. Use membrane filters which are 50 mm in diameter and having
• nominal pore size not greater than 0.45 µm.
• Pass the sample solution from filter paper .
• The filter is then rinsed and then the membrane is transferred
into the appropriate medium.
• Then the membrane is incubated for 14 days in the test
• medium.
2. Membrane filtration
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12. Finished product testing :
5. POTENCY TEST
Potency assay is one of the main methods used for assuring the
quality of vaccine which is based on the measurement of one or
several parameters that have been shown to be related directly
or indirectly with product efficacy. The main types of potency
tests performed for vaccine includes in-vitro titration of
antibodies a common lab test used to detect and measure
antibodies in the blood and in-vivo methods involving
immunization of small laboratory animals (e.g., mice, rats &
guinea pigs).
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13. ➢ IN VIVO POTENCY TEST
• Testing in-vivo potency/efficacy/assay or virus content of vaccine to
ensure that vaccinated vaccine recipient receives enough virus to
induce a protective immunity.
• Test animals are divided into two groups of 8-10 animals per group
and housed separately. One group of animals (adult mice, suckling
mice and guinea-pigs) is vaccinated while the other group remained
unvaccinated to act as control group
• Exposing vaccinated and unvaccinated animal with a virulent virus
strain of disease
In this method the animal is observed on daily basis.
Potency of test vaccine may be expressed as a percentage of potency of
standard vaccine. At least 90 percent of the vaccinated chickens
should survive the challenge and show no clinical signs of the
Newcastle disease should appear.
All unvaccinated control chickens should die of disease. Results of
biological assays is usually expressed in units of activity calibrated
against a reference standard.
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14. Invitro-potency assay :
Antibodies titration :A test used to detects the presence and measure
the amount of antibodies within a person’s blood.
Testing serum collected from vaccinated animal (serological test) for
level of antibodies titers. This Method is carried mostly
Titer level indicates that animal would be protected against disease
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15. An antibody titer refers to the highest dilution of a serum
sample that causes a positive test reaction:
Your text here
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16. Stability of vaccines
• Stability is the ability of a vaccine to retain its chemical, physical, microbiological and
biological properties within specified limits throughout its shelf-life.
• Shelf-life :The time period during which a vaccine may be stored and remain suitable for
use .
Factors Having a Strong Impact on Stability of Vaccines
• Purity
• Formulation
• Stabilizers
Human Serum Albumin (HAS), recombinant human Albumin
(rHA), Gelatine, Sugars, Sorbitol
Thiomersal
• Pharmaceutical form
• Lyophylized versus liquid
• Storage
• Frozen versus refrigerator
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17. Types of stability testing
1. Real time stability: checking long term stability under recommended
storage condition for the shelf life mentioned for the product.
2-8°C for Tetanus Vaccine
1. Accelerated Testing: Studies under exaggerated storage conditions.
15±2°C higher than the maximum temperature of storage at real
time storage of the product e.g. 25 ± 2°C
3. Stress studies: More severe conditions than those used for
accelerated studies. 35 ± 2°C
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18. Cold chain (no cold chain no Immunization)
System of storage and supply of vaccine at low temperature from the
manufacturer to the actual vaccination site :
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19. • Heat and Freeze sensitive vaccine
• Vaccines are grouped into 6
categories A,B,C,D,E,F
• A is more heat sensitive if heat is
increased it will deteriorate
• F is more freeze sensitive if it
become freeze it deteriorate.
• Heat sensitive should be place at
the lower portion of ILR (Ice lined
freezer)Like BCG ,OPV
• Freeze sensitive should be place at
upper portion of ILR(Ice lined
refrigerator).like Hepatitis B,TT
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20. • Light sensitive vaccine should be kept in amber colour vials (BCG and measles)
• Vaccine storage : +2 to +8 is normal cold chain temperature most of vaccine can stored upto 5 weeks in
this range of temperature.
OPV: Routine storage is 2-8 C but if long term storage needed then -15 to -25 C
Storage Devices used for storage of vaccine :
1. Ice lined refrigerator: Capacity 300 liters
2. Deep freezer : For Ice packs production
3. Cold chain box /vaccine carriers
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21. Shake test : If we suspect that the vaccine has frozen in past then we will conduct
shake test
Take a control vial and freeze it ,until completely solid (name it control vial) allow
this to become liquid again .
Now take the suspected vial ,shake both vials and observe for sedimentation.
If the suspected vial sediments slowly than control then use it .
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