Applications of Recombinant DNA

Technology in Biological Products like

Vaccines and Immune Products

 Recombinant DNA technology has rendered possible the

large- scale production of polypeptides present on the
surface of any pathogen.
 Contd..
• polypeptides, when purified from the producer organism can
then be used as ‘sub-unit’ vaccines.

• This method of vaccine production exhibits several

advantages:

(a) Production of a clinically safe product;

(b) Production of subunit vaccine in an unlimited supply;
(c) production of a defined product with less likely to cause
unexpected side effects.
 Types of Vaccines
(a)Subunit Vaccine:-

 large number of proteins are produced, isolated and

purified from the host cell and further processed to form
vaccine.
 Recombinant subunit vaccines made from a fragment of
protein (antigen) expressed in the laboratory using the
viral DNA.
 For Example, Vaccines for swine pseudo rabies, Foot &
Mouth disease & Hepatitis B has been Developed.
 Hepatitis B (HB) Vaccine
1. The hepatitis B virus (HBV) gene that codes for antigen
is inserted into baker’s yeast genome and then expresses
the antigen protein.

2. The antigen protein is harvested and purified to be used

for the vaccine.

3. This technique is also being used to explore a vaccine

against hepatitis C.
(b) Recombinant protein vaccination

 Upon infection, a pathogen produces proteins to elicit an

immune response from the infected body. The gene
encoding such protein is isolated to develop a rDNA which
is expressed in a heterologous expression system (e.g.,
bacterium, yeast, or insect).

 such as cholera vaccine, diphtheria toxoid, and tetanus

toxoid, are composed of protein/toxin antigens produced
in another host organism or purified from large amount of
pathogens.

 The vaccinated persons produce antibodies to the

protein/toxin antigen to protect themselves from diseases.
 Cholera Vaccine
• Cholera is an intestinal disease characterized by diarrhoea,
dehydration, abdominal pain and fever. It is caused by the
bacterium, Vibrio cholera.

• The currently used cholera vaccine is composed of phenol-

killed V. cholera. The immuno - protection, lasting for 3-6
months.

• By rDNA, it was possible to delete the DNA sequence

encoding A1 peptide and create a new strain of V. cholera.
This strain is non-pathogenic, and good candidate to serve
as an attenuated vaccine.
(c) Recombinant inactivated vaccines

 This vaccines consist of whole proteins extracted from the

disease agent or expressed from cloned genes in the
laboratory.

 Whole-cell expression systems include prokaryotic

(bacteria-based) systems such as E. coli, and eukaryotic
(mammalian, avian, insect, or yeast based) systems.

 Another type vaccines, called virus like particles

(VLPs), can be created when one or more cloned genes
that represent the structural proteins of a virus are
expressed simultaneously. These VLPs are immunogenic.
 Other Vaccines:
(a) Vaccinia Virus
• Vaccinia viruses is the vaccine that was used for eradication
of small pox.
• The cloned foreign genes (from pathogenic organism) can
be inserted into vaccinia virus genome for encoding
antigens which in turn produces antibodies against the
specific disease- causing agent.
• vector vaccine stimulates B-lymphocytes (to produce
antibodies) and T-lymphocytes (to kill virus infected cells).
• vaccinia virus can provide a high level of immuno
protection against pathogenic organisms.
• Vaccine for Malaria:
 The surface antigen of Plasmodium falciparum, one of
the 4 species of malaria, has been transformed to E. Coli
to produce amounts larger enough to develop a vaccine
against this form of malaria.

• Vaccine against Meningitis:

 A novel approach to develop a vaccine against
meningitis. They identified 350 proteins (potential
protective antigens) and entire sequence of genome
coding for these proteins in N. meningitides.
 All the 350 candidate antigens were expressed in E. coli,
purified and used to immunize individuals. A good
bactericidal antibody response was observed.
 Immune Products
• Production of monoclonal antibodies

 These are specific antibodies produced in large quantity ,

by fusing B-cells dervied from a single ancestral B-cell
with a tumour cell.

 These culture of B-cells are called monoclonal. These

cells are used to harvest single kind of antibodies called
as monoclonal antibodies.
• Lymphokines:
 These are proteins produced by lymphocytes. They
engage other lymphocytes to boost the immune response
to a foreign substance (antigen) and repel foreign
invasion or disease.

 lymphokines has been superseded by molecular cloning.

Large amounts of secreted lymphokines can be derived,
and monoclonal antibodies have provided valuable tools
with which to obtain homogeneous proteins.

 These have then been sequenced and cDNA libraries

prepared and cloning undertaken mostly in Escherichia
coli.
• Cancer Immunotherapy:
 Metastatic cancer can be regressed through
immunotherapy based on the adoptive transfer of gene-
engineered T-cells.

 Targeting of antigens expressed by tumors by the

successful use of gene engineering to retarget T-cells
before their transfer into the patient are mainly focused
on in this therapy.

 T-cells in cancer patients can be modified by

recombining the genes responsible for cancer-specific
antigens recognition, resistance to immunosuppression,
and extending survival and facilitating migration to
tumors.
 Other Immunity Products

 Adenosine deaminase (ADA) for treating some forms of

severe combined immunodeficiency (SCID)

 Adenoviruses in which critical virulence genes are

deleted have been used to express proteins from HIV19
and are being utilized in clinical trials for many other
pathogens such as the Ebola virus and malaria.
Prepared By:- Abdul Javeed

Roll.No:- 170716881005

B.Pharm 4th year

Section - A
JAZAKALLAHU
KHAIR