Recombinant Vaccines –

“A VACCINE PRODUCED FROM A CLONED GENE.”

A recombinant vaccine contains either a protein or a gene encoding a protein of

a pathogen origin that is immunogenic and critical to the pathogen function; the
vaccine is produced using recombinant DNA technology. The vaccines based on
recombinant proteins are also called subunit vaccines.

The logic of such vaccines, in simple terms, is as follows. Proteins are generally
immunogenic, and many of them are critical for the pathogenic organism. The
genes encoding such proteins can be identified and isolated from a pathogen
and expressed in E. coli or some other suitable host for a mass production of the
proteins.

It is created by utilising bacteria or yeast to produce large quantities of a

single viral or bacterial protein.This protein is then purified and injected into the
patient,and then the patient’s immune system makes antibodies to the
disease agent’s protein,protecting the patient from natural disease.

The concerned proteins are purified and mixed with suitable stabilizers and adjuvants after its
production, if required, and used for immunization. The different steps involved in the
development of a recombinant protein based vaccine may be simply summarised as follows.

(i) The first step is to identify a protein that is both immunogenic and critical for the
pathogen.
(ii) The gene encoding this protein is then identified and isolated.

(iii) The gene is integrated into a suitable expression vector and introduced into a suitable
host where it expresses the protein in large quantities.

(iv) The protein is then isolated and purified from the host cells.
(v) It is used for the preparation of vaccine.

The host organisms used for expression of immunogenic proteins to be used as vaccines may
be anyone of the following.

1. A genetically engineered microorganism, e.g., yeast for the expression of hepatitis B

surface antigen (HBsAg) used as vaccine against hepatitis B virus (approved for marketing in
India).

2. Cultured animal cells, e.g., HBsAg expressed in CHO (Chinese hamster ovary) cell line
and C-127 cell line.

3. Transgenic plants, e.g., HBsAg, HIV-l (human immunodeficiency virus-I) epitope (in
experimental stages).

4. Insect larvae; the gene is integrated into a bacculovirus genome, which is used to infect
insect larvae. Often a very high quantity of the recombinant protein is produced.
So far a large number of recombinant immunogenic proteins of pathogens have been
produced and evaluated. In general, a majority of such proteins are ineffective or only poorly
effective in immunization.

EXAMPLES OF RECOMBINANT VACCINE:

• GARDASIL: for HPV infection which causes genital warts and cervical cancer.
• LYME DISEASE VACCINE (recombinant OspA)
• MEASLES,MUMPS AND RUBELLA VACCINE LIVE
• HEPATITS B VACCINE:a preparation of HEP B surface antigen,derived either from
human plasma of carriers of HEP B or from cloning in yeast cells(hepatitis b virus).It
is the only licenced recombinant vaccine.

HEPATITIS
• Hepatitis is an inflammation of the liver.
• It is usually caused by viral infections, toxic agents or drugs but
may be an autoimmune response.
• It is characterised by jaundice, abdominal pain,liver enlargement
and sometimes fever.
• The different types of VIRAL hepatitis are A (formerly called
infectious hepatitis), B (serum hepatitis), C (formerly called non-A,
non-B hepatitis), D (delta hepatitis), E (a virus transmitted through
the faeces of an infected person), F, G, cryptogenic (Caused by a
virus as yet unidentified).
Characteristics of Hepatitis Viruses
Hepatitis Hepatitis Hepatitis Hepatitis Hepatitis
A Virus B Virus C Virus D Virus E Virus

Nucleic acid RNA DNA RNA * RNA

Serologic IgM anti- HBsAg Anti-HCV Anti- Anti-HEV
diagnosis HA HDV
Major Fecal–oral Blood Blood Needle Water
transmission
Incubation 15–45 40–180 20–120 30–180 14–60
period (days)
Epidemics Yes No No No Yes
Chronicity No Yes Yes Yes No
Liver cancer No Yes Yes Yes No
IgM anti-HAV = IgM antibody to hepatitis A virus; HBsAg = hepatitis B
surface antigen; anti-HCV = antibody to hepatitis C virus; anti-HDV =
antibody to hepatitis D virus; anti-HEV = antibody to hepatitis E virus.
*Incomplete RNA; requires presence of hepatitis B virus for replication.
HEPATITIS-B:
• It is a serious disease caused by a virus that attacks the liver. The
virus, which is called hepatitis B virus (HBV), can cause lifelong
infection, cirrhosis(scarring) of the liver, liver cancer, liver failure,
and death.
• HBV is a bloodborne pathogen, transmitted by percutaneous or
permucosal (e.g., sexual) exposure to infectious blood or body
fluids (e.g., semen or saliva).HBV circulates in high titers in the
blood and lower titers in other body fluids (e.g., semen, vaginal
fluid, or saliva), and is approximately 100 times more infectious
than HIV and 10 times more infectious than HCV.
Hepatitis B virus is an important cause of viral hepatitis. There is no specific
treatment for this disease.The incubation period for hepatitis B is relatively long; six
weeks to six months may elapse between exposure and the onset of clinical
symptoms. The prognosis following infection with hepatitis B virus is variable and
dependent on at least three factors: (1) Age — Infants and younger children usually
experience milder initial disease than older persons;1 (2) Dose of virus — The higher
the dose, the more likely acute icteric hepatitis B will result;1 and, (3) Severity of
associated underlying disease — Underlyingmalignancy or pre-existing hepatic
disease predisposes to increased morbidity and mortality.

• HBV is a 42nm, double-shelled deoxyribonucleic acid (DNA) virus of

the class Hepadnaviridae.
• The outer surface membrane contains hepatitis B surface
antigen(HBsAg), which also circulates in blood as 22-nm spherical
and tubular particles.
 • The inner core of the virus contains hepatitis B core antigen(HBcAg),
hepatitis B e antigen (HBeAg), a single molecule of partially double-
stranded DNA,and DNA-dependent DNA Polymerase.

HEPATITIS- B VACCINE:
Hepatitis B is given as a series of three intramuscular doses, 0,1 and 6
intramuscular injections for population at risk.

ENGERIX B (Glaxosmithkline) ,RECOMBIVAX HB(Merck & co.) and

GENEVAC B(Serum institute of India)are the three typical vaccines
developed for hepatitis b.

RECOMBIVAX HB:
RECOMBIVAX HB* Hepatitis B Vaccine (Recombinant) is a non-infectious subunit
viral vaccine derived from hepatitis B surface antigen (HBsAg) produced in yeast
cells. A portion of the hepatitis B virus gene, coding for HBsAg, is cloned into yeast,
and the vaccine for hepatitis B is produced from cultures of this recombinant yeast
strain according to methods developed in the Merck Research Laboratories.
The antigen is harvested and purified from fermentation cultures of a recombinant
strain of the yeast Saccharomyces cerevisiae containing the gene for the adw
subtype of HBsAg. The fermentation process involves growth of Saccharomyces
cerevisiae on a complex fermentation medium which consists of an extract of yeast,
soy peptone, dextrose, amino acids and mineral salts. The HBsAg protein is
released from the yeast cells by cell disruption and purified by a series of physical
and chemical methods. The purified protein is treated in phosphate buffer with
formaldehyde and then coprecipitated with alum (potassium aluminum sulfate) to
form bulk vaccine adjuvanted with amorphous aluminium hydroxyphosphate sulfate.
The vaccine contains no detectable yeast DNA but may contain not more than 1%
yeast protein. The vaccine produced by the Merck method has been shown to be
comparable to the plasma-derived vaccine in terms of animal potency (mouse,
monkey, and chimpanzee) and protective efficacy (chimpanzee and human).
The vaccine against hepatitis B, prepared from recombinant yeast cultures, is free of
association with human blood or blood products.Each lot of hepatitis B vaccine is
tested for sterility.
RECOMBIVAX HB is a sterile suspension for intramuscular injection. However, for
persons at risk of hemorrhage following intramuscular injection, the vaccine may be
administered subcutaneously.
RECOMBIVAX HB Hepatitis B Vaccine (Recombinant) is supplied in three
formulations.
Pediatric/Adolescent Formulation (Without Preservative), 10 mcg/mL: each 0.5
mL dose contains
5 mcg of hepatitis B surface antigen.
Adult Formulation (Without Preservative), 10 mcg/mL: each 1 mL dose contains
10 mcg of
hepatitis B surface antigen.
Dialysis Formulation (Without Preservative), 40 mcg/mL: each 1 mL dose
contains 40 mcg of
hepatitis B surface antigen.
All formulations contain approximately 0.5 mg of aluminum (provided as amorphous
aluminum
hydroxyphosphate sulfate, previously referred to as aluminum hydroxide) per mL of
vaccine. In each
formulation, hepatitis B surface antigen is adsorbed onto approximately 0.5 mg of
aluminum (provided as
amorphous aluminum hydroxyphosphate sulfate) per mL of vaccine. The vaccine is
of the adw subtype.
RECOMBIVAX HB is indicated for vaccination of persons at risk of infection from
hepatitis B virus
including all known subtypes. RECOMBIVAX HB Dialysis Formulation is indicated for
vaccination of adult
predialysis and dialysis patients against infection caused by all known subtypes of
hepatitis B virus.

CHRONIC AFFECT OF HEPATITIS B ACROSS THE WORLD:

Hepatitis B is one of the world's major health problems. By recent estimates, more than 2
billion people are infected with hepatitis B virus (HBV) globally. This includes 350 million
chronic carriers of the virus. The infection is supposed to be causally related to 1 to 2 million
deaths per year worldwide .In India, too, hepatitis B is a major public health problem. India
comes under the intermediate zone (2 to 7%) of HBV prevalence, the carrier rate being
approximately 5%.

Globally:

• 50 million new cases every year

• 350-400 million chronic carriers(75% in asia)

• 5,20000 death per year.

United states:

1. 1,40,000 to 3,20,000 new cases per year

2. 1.25 million chronic carriers(0.3% of adult population)

3. 4-5000 deaths per year.

ADVANTAGES OF RECOMBINANT VACCINE

• No chance of host becoming ill from the agent, since it is just a

single protein.

• It doesnot need an adjuvant.an adjuvant is an agent that stimulates

the immune system to find and react to the vaccine agent.some
adjuvants have been implicated in causing cancers in animals
overtime.

DISADVANTAGES OF RECOMBINANT VACCINE

• Isolated proteins may have a different three dimensional structure
than the protein in its normal context,and will induce antiobodies
that may not recognise the organism.

• Subunit vaccines often elicit weaker antibody responses than the

other classes of vaccine.