VACCINATION AND IT’S DELIVERY SYSTEM

Presenting By :-
Nikita R Mahajan
M-pharm-1st yr
[pharmaceutics]

INSTITUTE OF PHARMACEUTICAL EDUCATION

AND RESEARCH
BOARGAON {MEGHE} WARDHA
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CONTENT
• Introduction
• Definitions
• Advantages
• Disadvantages
• Types Of Vaccines
• How Vaccines Works
• Uptake Antigen
• Content Of Vaccine
• General Method Of Preparation Of Vaccine
• Quality Control Of Vaccine
• Single Shot Vaccine
• Delivery System Of Vaccines
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VACCINATION

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INTRODUCTION
 The word “vaccine” originates from the Latin
Variolae vaccinea (cowpox), which Edward Jenner
demonstrated in 1798 could prevent smallpox in
humans. Today the term ‘vaccine’ applies to all
biological preparations, produced from living
organisms, that enhance immunity against disease
and either prevent (prophylactic vaccines) or, in
some cases, treat disease (therapeutic vaccines).
Vaccines are administered in liquid form, either by
injection, by oral, or by intranasal routes.
 Ex. Polio , Hepatitis –A……etc.

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WHAT ARE VACCINES….?
 Vaccines are biological product which act by reinforcing
the immunological defense of the body against foreign
agencies or their toxins.
 The agent or product through which immunization is
active are called immunization agents.
 Active immunization is process of increasing resistance to
infection where by microorganism or product of their
activity act as a antigens and stimulate certain body cell
produce a antibodies with specific protective capacity.
 Passive immunization results in intermediate protecylon
of short duration is achaived by antibodies administration.
 Vaccines may be single component or mixed component
vaccines.

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ADVANTAGES
 A) Virosomes technology is approved by the FDA for use in humans,
and has a high .safety profile.

 B) Virosomes are biodegradable , biocompatible, and non-toxic.

 C) No disease-transmission risk .

 D) No auto immunogenic or anaphylaxis.

 E) Broadly applicable with almost all important drugs (anticancer

drugs , proteins, peptides, nucleic acids , antibiotics, fungicides)

 F) Enables drug delivery into the cytoplasm of target cell.

 G) Promotes fusion activity in the endolysosomal pathway .

 H) Protects drugs against degradation. 6

DISADVANTAGE

 Shelf-life is too short

 Scale up related problems
 Poor quality of raw material
 Pay-load is too slow
 Absence on any data on safety of these carrier systems on
chronic use.
 But in recent years several solutions have been worked
upon to overcome above

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TYPE OF VACCINS
 Vaccines are composed of either the entire disease-causing
microorganism or some of its components They may be
constructed in several ways.
Table no. 1

Sr . No TYPES OF VACCINS EXAMPLES

1 Live attenuated Measles , Mumps, Rubella
Etc
2 Inactivated Hepatitis-A, Influnza
,Pneumococcal.
3 Recombinant Hepatitis-B
4 Toxoid Tetanus, Diptheria.
5 Conjugate polysacchride protein Pnenococal ,
Meningococal , Influanze
–B ,
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Fig.No:-1
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 How do vaccines work?
 When inactivated or weakened disease-causing
microorganisms enter the body, they initiate an
immune response. This response mimics the body’s
natural response to infection. But unlike disease-
causing organisms, vaccines are made of components
that have limited ability, or are completely unable, to
cause disease.

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 Uptake of antigens
 The components of the disease-causing organisms or the
vaccine components that trigger the immune response are
known as “antigens”. These antigens trigger the production
of “antibodies” by the immune system. Antibodies bind to
corresponding antigens and induce their destruction by
other immune cells.
 The induced immune response to either a disease-causing
organism or to a vaccine configures the body’s immune
cells to be capable of quickly recognizing, reacting to, and
subduing the relevant disease-causing organism. When the
body’s immune system is subsequently exposed to a same
disease-causing organism, the immune system will contain
and eliminate the infection before it can cause harm to the
body.
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Fig.No:-3
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Antigens:-
 Antigens is an any substance which introduce parenterally in
body simulates the production of an antibody with it specifically
and an observable manner.
 Most antigens are either proteins or large polysacchrides .
 The complete antigen is able to induce antibody formation and
produce a specific and observable reaction with the antibody so
produced.
 The smallest unit of antigencity is known as the antigenic
determinant or epitope.
 The epitopic is that small area on the antigen usually consisting
of four or five amino acid or monosachride reduce possessing
specific chemical structure electrical charge and steric
configuration capable of sensitizing an immunocyte and of
reacting with its complementary site on the specific antibody or
T cell receptor.
 The combining area on the antibody molecules corresponding
to epitope is called paratope .
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Antibodies
 Antibodies are globulin proteins (A protein family with
compact globular form) therefore the term
immunoglobulin's ( Ig) for antibodies is used.
 Antibodies are made in response to an antigen and can
recognize and bind to the antigen .
 A bacterium or virus may have several epitopes that causes
the production of antibodies.
 Each antibody have at least two identical sites that binds to
epitopes known as antigen-binding site .
 The number of antigen-binding site on an antibody is
called the valence of that antibody. Most human antibodies
are bivalent.

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Figure no 4: Diagram of antibodies
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Immunoglobulin classes
 The simplest and abundant immunoglobulin's
are monomer but they can also assume some
difference in size and arrangement.
 The five classes of Is are designated IgG , IgM ,
IgA , IgD and IgE.
 Each class has a different role in the immune
response

Immunoglobulin class H chain

IgG g(gamma)
IgA a(alpha)
IgM m(mu)
IgD d(delta)
IgE e(epsilon)
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Antigen-Antibody reaction
 Antigens and antibodies combine with each other specifically
and in an observable manner.
 These reaction serve several purpose;
 In the body they form the basis of antibody mediate immunity in
infection diseases or of tissue injury in some type of
hypersensitive and autoimmune diseases.
 In the laboratory they help in the diagnosis of infections.
 In the identification of infectious agents and of noninfectious
antigens.
 These reaction can be used for the detection and quantization of
either antigen or antibodies.
 Antigen-antibody reaction in vitro are known as serological
reaction.

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 Both participate in the formation of agglutinate and
precipitates.
 Antigens and antibodies can combine in varying
proportion unlike chemical with fixed valence s . Both
antigens and antibodies are multivalent.
 Antibodies are generally bivalent though IgM molecule
may have five or ten combining site. Antigens may have
valance up to hundreds.

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Malaria vaccine
 : Malaria Vaccine develops antibodies against the
Plasmodiumfalciparum, the most aggressive type of
parasite that causes malaria. The parasite has two
main stages of development that take place into two
different organs. These are as follows.

Fig.No:-5 Action of malaria vaccines 21

 STEPS:
 Step 1-First, migration of the parasite transmitted by the
mosquito bite (sporozoite) to the liver.
 Step 2-Second, the sporozoite get transformed inside the
liver cell for generating hundreds of merozoites, which
then infect red blood cells.
 Therefore, proteins at the surface of the parasite
(sporozoites or merozoites) are constantly changing and
the ideal vaccine should be targeted through different
proteins from the various stages to improve the chance of
obtaining protection or reducing the symptoms related
mostly to the red blood cell infection.
 Vaccine is one of few designed for targeting the two forms
of the parasite: sporozoites (infecting liver cells) and
merozoites (infecting red blood cells). The vaccine uses
virosomes as a delivery platform.

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Component of vaccines

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 What does a vaccine contain?

In addition to the bulk antigen that goes into a vaccine,

vaccines are formulated (mixed) with other fluids (such as
water or saline), additives or preservatives, and sometimes
adjutants'. Collectively, these ingredients are known as the
excepients. These ensure the quality and potency of the
vaccine over its shelf-life. Vaccines are always formulated so
as to be both safe and immunogenic when injected into
humans. Vaccines are usually formulated as liquids, but
may be freeze-dried (lyophilized) for reconstitution
immediately prior to the time of injection.

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Preservatives

Preservatives Vaccines
Phenol Typhoid , Pneumococcal
Benzethonium chloride Anthrax
2-Phenoxyethanol Inactive Polio
Thimerosal Influenza

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Adjuvant
ADJUVANT VACCINES
Aluminum salt Hepatitis A& B, Tetanus,
HemopylusInfluanza B
,Pneumococcal, Tetanus,
,diphtheria.
Aluminum salt and mono Human Papilloma Virus
phospholipids

MF59[oil in water emulsion] H1N1 influenza

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General method for preparation of vaccines
 The seed lot system
• The starting stage of preparation of all microbial vaccines is the
isolation of suitable microbial strains.
• Microbial strain are mainly isolated from human infections and
in some cases have required elaborate laboratory manipulation
and selection.
• Once a suitable strain is available a sizeable culture is prepared
and distributed in large number of ampoules and then stored at
-70 degree or freeze dried. These culture is called ‘ seed lot’.
• The seed is then used to make one or more batches of vaccine
production .
• If it is found satisfactory then it is tested for efficacy and safety in
a clinical trials . Satisfactory result in a clinical trials validate the
seed lot and it is used for production of vaccines.
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Process of bacterial harvest

 The harvest is a complex mixture of bacterial cell and

metabolic product . The bacterial harvesting depend
on the nature of component that is required and ,may
involved one or more of the following procedure:
1. Killing
2. Separation
3. Fractionation
4. Detoxification
5. Adsorption
6. Conjugation

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Some examples
TAB Vaccine ( Typhoid-Paratyphoid A,B)
 It is a sterile suspension of Salmonella typhi and Salmonella paratyphi A
and B.
 TAB-C contain Salmonella paratyphi C in addition to TAB.
Preparation
 TAB vaccine is a mixed polyvalent vaccine and is prepared by mixing of
simple of vaccines of Salmonella typhi , S . paratyphi A and S. paratyphi
B.
 These strains are grow in acids digested agar medium and cultivated for
48 hr at 37 degree .
 Harvested with a sterile normal saline solution .
 Strain are diluted to form 3000 million organism/ml of Salmonella typhi
and 2250 million organism/ml of each S.paratyphi A and B.
 Above strain are killed by adding 0.1% formalin or by heat treatment.
 Bacterial strain are incubate at 37degree for 4 day for detoxification and
then tested for sterility.
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 bacterial strain are mixed together to contain 1000 million
organism of S. typhi and 750 million organism of each of S.
typhi A and B.
 The suspension is transferred to final sterile containers and
freeze dried.
 The sterility and abnormal toxicity is evaluate further.

Storage
 store in well closed container at temp 2-8 degree.
Dose
 Prophylactic 0.5 ml (sc)’ 2-4 injection 2-4 week interval.
 Booster dose give every 1-2 yr.
Uses
 TAB also mixed with tetanus and cholera vaccine.
 Prophylaxis of enteric infection. 30
MMR (measles-mumps-rubella) Vaccine
 Measles-Mumps-Rubella vaccines is a mixed preparation
containing suitable live attenuated of measles virus mumps virus
and rubella virus.
 Mortality due to measles is still high in countries due to
malnutrition.
 It is grown in a chick embryo cells.
 Single dose is injected (sc or i. m) in children older than 12
month to protect them from that three diseases.
BCG vaccines
 This is a suspension of living cell of strain of Mycobacterium
tuberculosis known as BCG vaccines.
 Originally it was given orally but to unreliable absorption from
gut the intracutaneous route is now preferred.
 The vaccine is prepared immediately before use by
reconstitution from the dried vaccine with suitable liquid.
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Quality control of vaccines
The quality control of vaccine is intended to provide
assurance of safety and efficacy . They are checked in two
ways;
 In-process : In this quality control is the control exercised
over starting material and intermediate.
The quality control of diphtheria and tetanus vaccine
required that the product are tested for the presence of
free toxins.
 Final product control : Final product control is the quality
control exercised by the monographs of a pharmacopoeia
of over product in their final container .

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 All vaccine are tested for;
1. Identity test
2. Potency test
3. Safety test
4. sterility assay
5. Free formalin test
6. Abnormal toxicity testing
7. Phenol concentration
8. Presence of aluminum and calcium

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Single shot vaccines

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The single shot vaccine concept
 The single shot vaccine is combination product of a prime
component antigen with an appropriate adjuvant and a
microsphere component that encapsulates antigen and provide
the booster immunization by delayed release of the antigen.
 Many aspects need to be taken into consideration when
developing such controlled released technology based vaccine .

Important determinants for single shot vaccine development

Biodegradable technology
Encapsulation Efficiency
Particle size distribution
Preservation of bio activity during formulation and released
Scalable production processes
Effects of combination with various adjuvant
Effects of different administration route
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Formulation and manufacturing of
single shot vaccines
 Important factors in the manufacture of a microsphere
based vaccine are high encapsulation efficiency and a
consistent particle production process. Several formulation
parameter play an important role in obtaining a robust
process. Below we discuss the process and equipment
used to manufacture several formulations.

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 Several factors are critical parameter for the formulation
of consist microsphere
FIRST., the size distribution of the microsphere can be
controlled by the shear a force applied during the
emulsification step in the bioreactor vessels. Factors that
has been identified to influence this shear force are the
mechanical stirring speed in the bioreactor vessels and
the viscosity of PEG solution which is determine by
concentration and molecular weight of the PEG.
SECOND, the presence of excipients in the starting
composition can influence the matrix density and
encapsulation efficiency to the microsphere product
either by direct effect on the microsphere formation or
on the protein characteristic . Finally polymerization
condition such as KPS concentration , pH, and
temperature ,can influence the strength of the form
hydro gel matrix
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 Controlling particle size during process scale-up.
 the dextran microsphere preparation method describe by
Steinke's etal., was initially perform on a 5-g scale
(containing 120mg of microsphere ),and used vortexing as
a means to emulsify the dex-HEMA phase in a continuous
PEG phase. However vortexing is not practically at a large
scale. Therefore we evaluate the feasibility of stirring, a
process that is relatively scale-up as a means of
emulsification ultimately at 500-g scale.

Fig no. microscopy picture of microsphere at a high stirring

speed of 700rpm (a) or a low stirring speed of 60 rpm (b).
 Nanocapsules for Single-Shot Vaccination

10000 Day 73
Day 103
Day 133
1000
Anti-HBsAg IgG (mIU/mL)

Day 163
Day 193
100

10

1
NC NC HBsAg-Alum
10µg (single-dose) 10µg (0,28) 10µg (0,28)

Sara Vicente et al. PLoS ONE 8(4): e62500, 2013.

 Transdermal delivery vaccine
 The skin is the largest and most accessible organ of the body . Vaccine
administration to skin offer many advantages include ease of access disease a
potential for generation of both systemic and mucosal immune response .
 The skin is a site of vaccine delivery it approaches to overcome these barriers are
cover included are formulation approaches such as liposome such as
electroporation , and technologies that creates micron size pores in the skin.
 The world health organization estimate that 32% of Hepatitis B Virus infections
,40% of Hepatitis C and 5% of Human Immunodeficiency Virus Infection in
developing country are attributable to unsafe injection practice .
 The development of needle free immunization method has thus becomes an
important goal in global health care .
 Dermal vaccination or transcutaneous immunization is a needle free method of
vaccine delivery which has the potential to reduced the risk of needle burn
disease ,access to vaccination by simplifying procedure (trained personal and use
of sterile equipments not required ) and assist in the implementation of multiple
boosting and multivalent vaccine regimes .
 Skin as a site for vaccine delivery the skin has multiple barrier properties to
minimize water loss from the body and prevents the permeation of
environmental contaminants into the body .
 These barrier can be considered as physical , enzymatic and immunological .

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 . They range from formulation approaches such as liposome's to
minimally invasive technology that creates channel in the skin such as
micro needle
 All method aim to over come the stratum conium barrier and target vaccines to
immune responsive cell such as langerhence cell.
 immunization by dermal route primary delivery method under investigation
and development.
a) liquid jet
b) Epidermal powder immunization.
c) Topical application of vaccines to the epidermis via
 hair follicles
 tap stripping to remove the stratum corneum
 Thermal or radio-wave mediated ablation of the stratum cornea
 Colloidal carriers such as micro emulsion and liposome's increase dermal
absorption
 Low frequency ultrasound as an adjuvant and to increase skin penetration
 topically applied adjuvant to induce a potent immune responses
 Electoporation of the stratum cornea
 Shallow micro niddles that penetrate into the epidermis
Application transdermal
 application of liquid-jet injector focused on delivery
of macromolecules that do not passively permeate the
skin.
 For administration of insulin and human growth
hormone
Mucosal delivery vaccines

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Mucosal delivery vaccines

Systemic-
Needle-free
mucosal
vaccination
response

Natural Easier
entrance production
Mucosal
vaccines
 Mucosal vaccines

Challeng Challenges Nanotechnology Solution

Enzymes Protection

Penetration
Barriers
Targeting

Ag release Sustained

Stimulation Adjuvants
REFERENCE
 Drug And Pharmaceutical Science ; Novel Drug
Delivery System Second Edition By Tie W. Chien
Published By Marcel Dekker New York; Hongkong
1992 Pg No. 197;301.
 N.K.Jain , Controlled and Novel Drug Delivery , CBS
Publisher & Distributer , New Delhi , First Edition
1997.Pg.No;-503 507
 K.R Kuchekar , Microbiology And Immunology, By
Nirali Prakashan New Delhi. Pg No 57-62.

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