Production of Interferon, Hepatitis Vaccines & Insulin Hormones

1. Application of rDNA
in the Production of Interferon, Vaccine Hepatitis-B, Hormones- Insulin
2 July 2021 Abhijit Debnath BP605T and Biotech Unit-1 1
CO1.1
Noida Institute of Engineering and Technology
(Pharmacy Institute) Greater Noida
Abhijit Debnath
Asst. Professor
NIET (Pharmacy Institute)
Unit: 2
Subject Name: Biotechnology
(BP605T)
Course Details
(B. Pharm 6th Sem)

2. Application of
rDNA in the
Production of
2 July 2021 Abhijit Debnath BP605T and Biotech Unit-1 2
 Interferon
 Vaccine Hepatitis-B
 Hormones- Insulin
CO1.1
Noida Institute of Engineering and Technology
(Pharmacy Institute) Greater Noida

3. • Interferons (IFNs, /ˌɪntərˈfɪərɒn) are a group of signaling proteins made
and released by host cells in response to the presence of several viruses.
In a typical scenario, a virus-infected cell will release interferons causing
nearby cells to heighten their anti-viral defenses.
• IFNs belong to the large class of proteins known as cytokines, molecules
used for communication between cells to trigger the protective defenses
of the immune system that help eradicate pathogens
• Naturally occurring proteins and glycoproteins Secreted by eukaryotic
cells in response to viral infections, tumors, and other biological
inducers.
• Strucurally, they are part of the helical cytokine family which are
characterized by an amino acid chain that is 145-166 amino acids long.
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INTERFERONS (CO2.2)

4. • IFNs also have various other functions:
• they activate immune cells, such as natural killer cells and macrophages;
• they increase host defenses by up-regulating antigen presentation by virtue of increasing the expression of
major histocompatibility complex (MHC) antigens.
• Certain symptoms of infections, such as fever, muscle pain and "flu-like symptoms", are also caused by the
production of IFNs and other cytokines.
• More than twenty distinct IFN genes and proteins have been identified in animals, including humans.
• They are typically divided among three classes: Type I IFN, Type II IFN, and Type III IFN.
• IFNs belonging to all three classes are important for fighting viral infections and for the regulation of the immune
system.
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INTERFERONS (CO2.2)

5. 2 July 2021 Abhijit Debnath BP605T and Biotech Unit-2 5
INTERFERONS (CO2.2)

6. 2 July 2021 Abhijit Debnath BP605T and Biotech Unit-2 6
INTERFERON (CO2.2)

7.  Interferon therapy
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INTERFERONS (CO2.2)
Diseases
 Interferon beta-1a and interferon beta-1b are used to treat and control multiple sclerosis, an autoimmune
disorder. This treatment may help in reducing attacks in relapsing-remitting multiple sclerosis and slowing
disease progression and activity in secondary progressive multiple sclerosis.
 Interferon therapy is used (in combination with chemotherapy and radiation) as a treatment for some
cancers. This treatment can be used in hematological malignancy, such as in leukemia and lymphomas
including hairy cell leukemia, chronic myeloid leukemia, nodular lymphoma, and cutaneous T-cell lymphoma.
 Patients with recurrent melanomas receive recombinant IFN-α2b.
 Both hepatitis B and hepatitis C are treated with IFN-α, often in combination with other antiviral drugs. Some
of those treated with interferon have a sustained virological response and can eliminate hepatitis virus.

8.  Interferon therapy
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INTERFERONS (CO2.2)
Drug formulations
 Several different types of interferons are approved for
use in humans.
 One was first approved for medical use in 1986.
 In January 2001, the Food and Drug Administration
(FDA) approved the use of PEGylated interferon-alpha in
the USA
 Interferon-containing regimens may also include
protease inhibitors such as boceprevir and telaprevir.
 There are also interferon-inducing drugs, notably
tilorone that is shown to be effective against Ebola
virus.

9.  Interferon Production by rDNA
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INTERFERONS (CO2.2)

10.  Constitutional Toxicities
 Hematologic Toxicities
 Organ Toxicities
 Neuropsychiatric Toxicities
 Endocrine and Metabolic Toxicities
 Potential Drug Interactions
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CLINICAL TOXICITY OF INTERFERON
ADMINISTRATION (CO2.2)

11.  Hepatitis B is caused by the hepatitis B virus
(HBV), an enveloped virus containing a partially
double stranded, circular DNA genome.
 Sign & Symptoms:
11
2 July 2021 Abhijit Debnath BP605T and Biotech Unit-2
HEPATITIS B VACCINE (CO2.2)
 Fever
 Fatigue
 Loss of appetite
 Nausea
 Vomiting
 Abdominal pain
 Dark urine
 Clay-colored bowel movements
 Joint pain
 Jaundice
 Hepatomegaly
 Symptoms begin an average of 3 months (range: 2–5months) after exposure to HBV.
 Symptoms typically last for several weeks but can persist for up to 6 months.

12.  Its an ancient disease first described in 5th centuryB.C.
 Earliest recognized blood-borne outbreak of hepatitis was in Germany in
1883 after receiving smallpox vaccine.
 In 1947 MacCalum and Bauer introduce the term Hepatitis A for infectious
and Hepatitis B for serum hepatitis
 This terminology was adopted by WHO in 1973
 The term hepatitis describes inflammation of the liver.
 Hepatitis may be caused by alcohols, drugs, autoimmune diseases,
metabolic diseases, and viruses. Viral infections accounts for more than
half the cases of acute hepatitis.
 Viral hepatitis is a systemic infection affecting the liver predominately with
primary inflammation of the liver by any one of a heterogeneous group of
hepatotropic viruses.
o
5
2 July 2021 Abhijit Debnath BP605T and Biotech Unit-2 12
HEPATITIS B VACCINE (CO2.2)
 Hepatitis A (HAV)(1973)
 Hepatitis B (HBV) (1970)
 Hepatitis C (HCV) (1988)
 Hepatitis D (HDV) (1977)
 Hepatitis E (HEV) (1983)
 Hepatitis F – Not separate entity –
Mutant of BVirus.
 Hepatitis G (HGV) (1995)
TYPES OF HEPATITIS VIRUSES

13. 13
2 July 2021 Abhijit Debnath BP605T and Biotech Unit-2
HEPATITIS B VACCINE (CO2.2)

14. Insulin is a hormone produced by the beta cells of pancreas that permits
glucose to enter cells and help the body use glucose for energy. Insulin control
the amount of glucose in the blood.
One of the more important uses of inhaled insulin will be in individuals with
type 2 diabetes, especially those who fail combination oral therapy. Regimens
using pre-prandial inhaled insulin along with one injection of basal insulin have
been shown to be comparable to conventional insulin injection regimens
(mixed regular/neutral protamine Hagedorn insulin) in subjects with type 2
diabetes. However, it has also been reported that patients with either type 1 or
type 2 diabetes receiving inhaled insulin have reported enhanced overall
satisfaction, quality of life, and acceptance of intensive insulin therapy.
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INSULIN (CO2.2)

15. PROCESS OF PRODUCING INSULIN USING
RECOMBINANT DNA TECHNOLOGY (CO2.2)
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16. • The human gene is isolated. The mRNA is taken from the cell of islet of Langerhans.
 Messenger RNA is a molecule of RNA that encodes a chemical
• "blueprint" for a protein product.
 The isolated gene contains the code of the human DNA for the production of
insulin.
• The plasmid DNA of the bacterial cell is taken out of the cell. NOTE: Escherichia coli E.
Coli) bacteria is widely used in producing insulin but yeast may also be used.
FIRST STEP (PREPARING) (CO2.2)
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17. The plasmid DNA of the bacteria is cut out producing plasmid
ring which is an empty segment of the DNA.
 A Restriction Enzyme is an enzyme that cuts DNA at specific
recognition nucleotide sequences known as restriction sites.
 A segment of DNA known as sticky ends.
SECOND STEP (CUTTING) (CO2.2)
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18. • With the plasmid ring open, the gene
obtained from human cell that contains the
code of protein responsible for the
production of insulin is inserted into the
plasmid ring and the ring is closed.
• The human insulin gene is now combined
with the bacterial DNA plasmid.
Mix the recombinant plasmid
with bacteria.
THIRD STEP (COMBINING) (CO2.2)
2 July 2021 Abhijit Debnath BP605T and Biotech Unit-2 18

19. Transformed: resulting DNA is inserted back
to the bacteria.
FOURTH STEP (INSERTING) (CO2.2)
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20. The cells need nutrients in order to grow,
divide, and live. While they live, the
bacterial cell processes turn on the gene
for human insulin and the insulin is
produced in the cell. When the bacterial
cells reproduce by dividing, the human
insulin gene is also reproduced in the
newly created cells.
Production of Insulin (CO2.2)
2 July 2021 Abhijit Debnath BP605T and Biotech Unit-2 20

21. 2 July 2021 Abhijit Debnath BP605T and Biotech Unit-1 21
 Making of Insulin
Production of Insulin (CO2.2)