Quality Assurance

of Vaccines
by Shahid Ali Khan MIIPS
Contens

▶ Introduction
▶ Stability of Vaccines
▶ Potency Testing of Vaccines
▶ Post Market Surveillance of Vaccines
Introduction

▶ Vaccine
˝Vaccine is a suspension of weakened, killed, or
fragmented microorganisms or toxins or of antibodies or
lymphocytes
that is administered primarily to prevent disease
.˝ OR
˝Vaccines by definition are biological agents that
elicit an
immune response to a specific antigen derived from
an
infectious disease-causing pathogen.˝
▶ First Vaccine
Edward Jenner developed the first vaccine in 1796 using cowpox
to inoculate against smallpox.
Mechanism of Action of Vaccines

▶ Vaccine can confer

1.Active Immunity against a specific harmful agent by stimulating the
immune system to attack the agent through antibodies production.
2. Passive Immunity by providing antibodies or lymphocytes already made by
an animal or human donor.
Types of vaccines

▶ Four types of vaccines are currently available:

1.Live attenuated vaccines:
These contain a version of the living virus that has been weakened so that
it does not cause serious disease in people with healthy immune systems.
E.g. influenza vaccine
2.Inactivated vaccines:
These contain pathogens that are inactivated by heat or
chemical treatment.
E.g. hepatitis A vaccine.
Types of vaccines

3.Subunit, recombinant, polysaccharide, and conjugate vaccines:

These are biosynthetic vaccines that contain man-made substances which are
very similar to the virus or bacteria.
E.g. hepatitis B vaccine
4.Toxoid vaccines:
These are made from selected toxins that have been sufficiently attenuated and
are able to induce a humoral immune response. These toxins produce many of the
symptoms of the disease.
For example, diphtheria and tetanus vaccines
Quality Assurance of Vaccines

▶ Vaccines require special quality control and quality

assurance mechanisms because:
 These are derived from living organisms with a molecular composition
too complex to be defined by physical or chemical means.
 The inherent variability of living organisms and the potential for
contamination of materials derived from them.
Quality Assurance of Vaccines

▶ The manufacturer has the primary legal responsibility for the safety,
quality, and efficacy of the vaccines.
▶ The National Regulatory Authorities (NRAs),in the countries where
vaccines are manufactured, play a critical role in assuring product
quality.
▶ WHO first adopted recommendations for the national control of vaccines
and sera in 1981.
Quality Control Tests For Vaccines

Quality control tests are performed

on:
▶ Starting or raw material
▶ Intermediate or in process tests
▶ On final product
Quality Control Tests For Vaccines

▶ Safety Testing
▶ Potency Testing
▶ Stability Testing
▶ Post-Marketing
Surveillance
▶ Immunogenicity
▶ Bioassay
1)Safety Testing

▶ The animal safety tests are performed to detect vaccine toxicity

because residual vaccine toxicity has the potential to cause adverse
reactions.
▶ The animal body weight change test is the most commonly used test
to evaluate the toxicity of vaccines.
2) Potency Testing
▶ Potency denotes the amount of drug required to produce desired effect.
▶ Potency testing is one of the main methods used for assuring the quality of
vaccine.
▶ This test is based on the measurement of one or several parameters that are
related directly or indirectly with product efficacy.
▶ For live, attenuated vaccines in-vitro potency assay is used while it is not
commonly used for inactivated vaccines.
IMPORTANCE OF POTENCY TESTING

The potency testing provide information about

▶ The mean potency value for a lot
▶ Manufacturing variability and Assay uncertainty of an individual vial
from the same vaccine lot.
▶ The characteristics of the lot of vaccine from which the vial has
taken (mean potency) but we can never know the actual potency
in the individual vial that was used in clinical trials.
CLINICALLY RELEVANT PARAMETRS FOR
POTENCY TESTING

▶ L.L (Lower limit):

the lowest dose at which the vaccine is going to be
effective.
▶ U.L (Upper limit):
the highest dose at which the vaccine is going to be safe.
These are normally specifications.
Potency testing Methods

(A) The Traditional Vaccine Potency Testing Method

(B)The Alternative Vaccine Potency Testing
Methods
1. Experimental animal substitution method
2. Serological substitution method
(A) The Traditional Vaccine Potency Testing Method

▶ To evaluate the quality of a vaccine, the potency test of the final product
must strictly enforce the standards of efficacy established by national
regulatory authorities.
▶ The universal method for testing the efficacy of vaccines is the
animal challenge test.
▶ Inoculate the animal with the recommended immunological dose
▶ To the prescribe day after the optimal immune response period, attack
the immunized animal and the control animal at the same time with
pathogens which are capable of causing the animal's pathogenesis, to
measure the ability of the vaccine, and to reduce the incidence of the
disease after a certain time according to the protection of the immunized
animal.
(B) The Alternative Vaccine Potency Testing Methods

1. Experimental Animal Substitution Method:

This method involve the use of other animals with similar clinical symptoms
to replace the original animals, and indirectly test the efficacy of the
animal vaccine.
2. Serological Substitution Method:

The efficacy of vaccine is assessed by detecting the antibody titer level

after immunization. It involves viruses neutralizing experiments and all kinds
of ELISA (Enzyme-Linked Immunosorbent assay, commonly used analytical
biochemistry assay)
3) Stability Testing

▶ ˝ Stability is the ability of a vaccine to retain its chemical, physical,

microbiological and biological properties within specified limits
throughout its shelf-life.˝
▶ For stability testing of vaccines the guidelines provided by ˝The
International Conference on Harmonisation (ICH)˝ for stability testing of
biological products are used.
 3)Stability
Testing
• ▶ The stability of vaccines has a major impact on the success of
immunization program worldwide.
•▶ to assure vaccine quality, WHO has
 acknowledged the importance of clearly defining the stability
• characteristics of a vaccine and
 emphasizes the role of national regulatory authorities in overall vaccine
evaluation.
Stable vaccine

▶ A vaccine with a very long shelf life.

OR
▶ A vaccine lot that complies with release specifications throughout
shelf life.
OR
▶ Vaccine that complies with the principles of the initial license and
technical, non-clinical and clinical specifications throughout its life
cycle.
Types of Vaccines Stability
testing
I. Real-time Stability Testing
II. Accelerated Stability Testing
III. Extreme Conditions Stability
Testing
IV. Thermal Stability Testing
I. Real-time Stability
Testing

▶ This test involve the observation and study of changes in

 the physical, chemical and biological characteristics of vaccine
 efficacy of the vaccine
over a period of time under the expected conditions of use and
preservation.
ii. Accelerated Stability
Testing

▶ This test involve the study of the rate of change in physical, chemical
and biological characteristics of a vaccine that is placed at sequence
temperature including above the actual preservation temperature.
▶ This study provides supporting data on the shelf-life or release criteria
for vaccines.
iii. Extreme Conditions (stress) Stability Testing

▶ The vaccine is kept under extreme environment conditions, such as,

light and extreme temperatures.
▶ The main purpose is to determine
 Packaging and labeling requirements,
 shipping,
 preservation and use considerations
 measures to be taken once exposed to extreme conditions.
iv. Thermal Stability Testing

▶ The vaccine is kept at higher storage temperature after a certain period

of time, to observe the
 main quality standards of compound situation and
 change ratio, which is an important component of the vaccine to
approve and release standard
▶ The main purpose is
 to assess whether a batch of vaccine stable enough and
 to determine whether a product batch and batch quality are uniform
Conditions for stability
testing
Type of testing Temperature Duration
Real-time 2-8˚C 4 years
Accelerated 20-25±2˚C 6months
Stress 35±2˚C 1-2months
Acceptance criteria for
stability testing
In general the significant change is define as
▶ 5% change in assay from the initial value or failure to meet
the acceptance criteria by biological or immunological
parameters
▶ Any degradation product (impurity) exceeding the specified
limit
▶ Failure to meet the specifications for physical and chemical
parameters
Stability of combined vaccines

▶ Determination of the shelf-life should be based on the shortest shelf-

life component
▶ Data obtained for the monovalent vaccines should support stability
of combined vaccine
▶ Stability of combined vaccines should not be based on the
extrapolation
of the stability data of the individual components alone.
Importance of stability testing

Stability studies:
 Determine shelf-life and storage conditions to support
licensing.
 Monitor vaccine stability in the post-licensure period.
 Support manufacturing changes
4) Post Market Surveillance
(PMS)of Vaccines

▶ “Post-market surveillance refers to all the processes that are carried out
to continuously track/monitor quality, safety and efficacy of vaccines in
the market after registration.”
▶ It helps to detect:
 Rare ADRS
 Drug interactions
 New uses for drugs
▶ Types of PMS:
1) Active(proactive) PMS
2) Reactive PMS
Active
Reactive PMS
PMS
▶ Active PMS refers to coordinated ▶ Reactive PMS refers to: follow up
surveys, sampling, analysis, on complaints from spontaneous
evaluation and assessment of reporting.
regulatory requirements in relation to
labeling and storage etc.
Scheme For
PMS
Need For
PMS
▶ Vaccines have risk that may include rare serious adverse events
not detected.
▶ Therefore post-market surveillance is essential
 To assure product safety.
 Detect manufacturing problems
 To check performance in different user populations
 To check product sustainability
Approaches for PMS of Vaccines

Two approaches are used for PMS of

vaccines:
A) Passive Approach
B) Active Approach
A) Passive Approach

▶ This approach uses Vaccine Adverse Event Reporting System (VAERS) which is
 A National System for passive surveillance of adverse events after
vaccination established in 1990.
 Jointly managed by FDA and CDC(Centers for Disease Control)
▶ The purpose of VAERS is to detect possible signals of adverse events
associated with vaccines.
▶ VAERS collects and analyzes information from reports of adverse
events (possible side effects) that occur after the administration of US
licensed vaccines.
▶ Reports are welcome from all concerned individuals: patients,
parents, health
care providers, pharmacists and vaccine manufacturers.
VAERS: Advantages

▶ National in scope, covers diverse

population
▶ Able to detect rare adverse events
▶ Rapid detection of possible signals
▶ Can assess adverse events by lot
VAERS: Limitations

▶ Reported diagnosis not verified

▶ Lack of consistent diagnostic criteria
▶ Under-reporting
▶ No unvaccinated control group
▶ Usually not possible to assess whether vaccine caused the
reported adverse event
B) Active Approach

▶ This approach uses CDC’s Vaccine Safety Datalink (VSD).

▶ It is a collaborative project between the Centers for Disease Control
and Prevention (CDC) and 9 health care organizations.
▶ Established in 1990.
▶ VSD is a vital resource informing policy makers and the public about
the safety of vaccines used in the United States.
▶ Large linked databases are used to identify and evaluate adverse
events in over 9 million individuals annually.
Advantages of
VSD
▶ VSD generates rapid, important safety assessments for both
routine vaccinations and emergency vaccination campaigns.
▶ Allows calculations of background rates of various conditions of
interest.
Limitations of VSD

▶ Sample size may be inadequate for very rare adverse

events
▶ Unvaccinated population may be small
▶ Variable accuracy of coded data used for studies
Importance of
PMS
▶ It play an important role to discover an undesirable effect that
might present at risk.
▶ It provide additional information on the benefit and risk of
vaccines.
Limitations of
PMS
▶ No fixed duration
▶ No fixed patient population
5) Immunogenicity
• ▶ DNA microarray analysis is used to translate vaccine quality,
immunogenicity, and reactogenicity, into gene expression profile data.
•▶ This method is expected to be
 Informative
 Rapid
 highly sensitive
Method For Testing Immunogenicity

▶ Take 8-week-old male rats(3-6), weighing 180–220 g.

▶ Intraperitoneally administered rats with 5 mL of vaccine or
physiological saline (SA).
▶ Sacrificed vaccinated rats to obtain whole lung, kidney, brain, and
the
lateral left lobe of the liver on day 1, 2, 3, and 4 after administration.
▶ Immediately freeze the tissues in liquid nitrogen for storage.
▶ Purify the DNA from the tissues and subject it to DNA microarray
analysis.
6) Bioassay

▶ A bioassay is an analytical method to determine concentration

or potency of a vaccine by its effect on living cells or tissues.
▶ Bioassay is performed on
 Polio vaccine
 Diphtheria vaccine
 BCG vaccine (etc)
?
References

▶https://www.who.int/biologicals/vaccines/regulation_and_quality_control_
vaccines/en/
▶https://www.britannica.com/science/vaccine

▶https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4257027/

▶https://www.creative-biolabs.com/vaccine/the-potency-tests-of-
vaccines.htm
▶https://pubmed.ncbi.nlm.nih.gov/25108215/

▶https://www.fda.gov/vaccines-blood-biologics/report-problem-center-
biologics-evaluation-research/vaccine-adverse-events
Thank you