The document summarizes several current biotech products including interferon, interleukin, human insulin, recombinant vaccines, monoclonal antibodies, trastuzumab, and follicle stimulating hormone. It describes how each product is produced through recombinant DNA technology, including gene isolation, vector insertion, cell culture expression, and purification processes. Key details like storage conditions and marketed preparations are also provided for several of the biotech products.

1. Short study of current
Biotech products
Presented by
Kazi Ornob
Fateh Al Mujib
Imrul Hasan Mim

2.  Biotech Products
 Interferon
 Interleukin
 Human Insulin
 Recombinant vaccines
 Monoclonal antibody
 Trastuzumab
 Follicle Stimulating Hormone
Contents

3. Biotech
products
 Biotech products broadly refers to
biopharmaceuticals drugs generated through
researches in cell biology, genetics and recombinant
DNA technology e.g.
Insulin
Antibiotics
Monoclonal antibodies
Vaccines
Interferon and interleukins
Follicle stimulating hormones etc.
 These products are essential for human body and
when we lack one of these elements it becomes vital
to be taken from outside the reason why studying the
manufacture of these products is so important.

4. Interferon
 An antiviral substance & is the first line of
defense against viral attacks.
 Term ‘interferon’ originated from the
‘interference’ of this molecule on virus
replication.
 Interferons are a family of host coded
proteins produced by cells on induction
by viral or non-viral inducers.
 Interferons by itself has no direct action
on viruses but it acts on other cells of the
same species, rendering them refractory
to viral infection.

5. The complementary DNA(cDNA) was synthesized
from mRNA of a specific interferon.
Inserted to a vector(plasmid) which is introduced
into E.coli or other cells.
IFN can be isolated from culture medium.
This is the basic mechanism of producing
recombinant IFNs.
Productionof
Recombinant
Interferons

6. Storage
Condition
 Interferons are inactivated by
proteolytic enzymes but not by
nucleases or lipases.
 They resist heating at 56-60°c for
30-60mins & stable over a range
of pH 2-10, except gamma IFN
which is liable at pH 2.
 Interferons are stored at 2 – 8°c

7. Marketed Preparation
Brand name Type Use
Alferon N Human leukocyte– derived
interferon alfa- n3
Genital and perianal warts
Roferon-A Recombinant interferon
alfa-2a
Hairy cell leukemia, AIDS
Intron- A Recombinant interferon
alfa-2b
Hairy cell leukemia, AIDS
Avonex, Rebif Recombinant interferon
beta-1a
Multiple sclerosis
Betaseron Recombinant interferon
beta-1b
Multiple sclerosis

8. Interleukins
These are a large group of cytokines
produced mainly by T-cells, although some
are also produced by mononuclear
phagocytes or by tissue cells.
There are currently 35 well known
interleukins, however, there are there are
many more to be found and characterized.
For example, IL-1, IL-2, IL-3, IL-6 etc.

9. Productionof
Interleukins
A gene coded for polypeptide which
possesses interleukin -2 activity is isolated .
Inserted into a suitable vector
The vector is transferred into the bacteria .
After that bacteria is grown in a culture
media .
Isolation and purification

10. Storage
Recombinant human IL is shipped at
room temperature. Upon receipt it
should be stored at -20°C
Note: Avoid repeated freeze-thaw cycles.

11. Marketed Preparation
Brand Name Type Use
ALDESLEUKIN Recombinant human IL- 2
(rIL-2)
Chronic hepatitis C, and
Chronic hepatitis B
SIGOSIX IL-6 Multiple sclerosis
OPRELVEKIN Recombinant human IL- 11 Hairy cell leukemia, AIDS
MUPLESTIM IL-3 Hairy cell leukemia, AIDS

12.  Earliest use of recombinant technology
 Modified E.coli cells to produce insulin; performed by
Genentech in 1978
 Prior, bovine and porcine insulin used but induced
immunogenic reactions
 To overcome this problems, researchers inserted
human insulin genes into a suitable vector (E.coli)
HumanInsulin

13. Producing
Recombinant
Insulin
 First, synthesized genes for the two insulin A & B
chains were inserted into plasmids along with a strong
lacZ promoter.
 The genes were inserted in such a way that the insulin
& B-galactosidase residues would be separated by a
methionine residue. This is so that the insulin A & B
chains can be separated easily by adding cyanogen
bromide.
 The vector was then transformed into E.coli cells.
 Once inside the bacteria, the genes were "switched-
on" by the bacteria to translate the code into either
the "A" chain or the "B" chain proteins found in insulin
 The purified insulin A and B chains were then attached
to each other by di-sulphide bond formation under
laboratory conditions

14. Storage
Insulin should be stored in the
refrigerator. If refrigeration is not
possible, it can be kept at room
temperature [15-25 degrees C] for
28 days

15. Recombinant
Vaccines
• Two types:
(i) Recombinant protein vaccines: This is
based on production of recombinant DNA
which is expressed to release the specific
protein used in vaccine preparation
(ii) DNA vaccines: Here the gene encoding for
immunogenic protein is isolated and used to
produce recombinant DNA which acts as vaccine
to be injected into the individual.

16. Productionof
Recombinant
proteinvaccines
 A pathogen produces its proteins in the
body which elicit an immune response from
the infected body.
 The gene encoding such a protein is isolated
from the causative organism
 This DNA is expressed in another host
organism, like genetically engineered
microbes; animal cells; plant cells; insect
larvae etc. resulting in the release of
appropriate proteins.
 These when injected into the body, causes
immunogenic response against the
corresponding disease thus, providing
immunity.

17. DNAvaccines
 Refers to the recombinant vaccines in which the
DNA is used as a vaccine.
 The gene responsible for the immunogenic protein
is identified, isolated and cloned with corresponding
expression vector.
 Upon introduction into the individuals to be
immunized, it produces a recombinant DNA
 This DNA when expressed triggers an immune
response and the person becomes successfully
vaccinated.
 The mode of delivery of DNA vaccines include:
direct injection into muscle; use of vectors like
adenovirus, retrovirus etc. in-vitro transfer of the
gene into autologous cells and re-implantation of the
same and particle gun delivery of the DNA.

18. Productionof
DNAvaccines:
 In certain cases, the responsible gene is
integrated into live vectors which are
introduced into individuals as vaccines.
 This is known as live recombinant
vaccines.
E.g: Vaccinia virus. Live Vaccinia virus
vaccine (VV vaccine) with genes
corresponding to several diseases, when
introduced into the body elicit an immune
response but does not actually cause the
diseases.

19. Monoclonal
antibody
Antibodies that are made by
identical immune cells that are
clones of unique parent cell, which
have monovalent affinity, in that
they bind to the same epitope.

20. Hybridoma
Production
The production of hybridoma that
continuously secret the useful
monoclonal antibody requires several
steps, namely-
1.Immunization
2.Cell fusion
3. Selection
4.screening
5.Cloning
6.Characterization and storage

21. Trastuzumab
 A monoclonal antibody
 It is sold under the brand name Herceptin.
 Used to treat breast cancer.
 Herceptin was developed using rats, mice,
hamsters and macaques.
 It was the first monoclonal antibody to treat
cancer successfully.
Mechanism of actions:
 Herceptin is designed to attach the protein HER2.
 HER2 makes cancer cells replicate faster.
 Herceptin prevents HER2 from working and
 Causes the cancer cells to die in patients whose
cancer has high levels of HER2 protein.

22. Follicle
Stimulating
Hormone
Follicle-stimulating hormone (FSH) is a
gonadotropin, a glycoprotein polypeptide
hormone. FSH is synthesized and secreted by
the gonadotropic cells of the anterior
pituitary gland, and regulates the
development, growth, pubertal maturation,
and reproductive processes of the body.

23. Preparation of recombinant human follicle-
stimulating hormone.
 An aliquot from the selected clone of Chinese
hamster ovary cells is first grown in tissue culture
flasks,
 then sub-cultured into roller bottles,
 and allowed to expand for up to 36 days.
 The cells are then mixed with a suspension of
micro-carrier beads.
 and transferred to a bioreactor for an average
duration of 34 days.
 The harvested crude FSH is stored at 48 °C until
purification.
 The final product is released after extensive
quality control testing over a period of 7 weeks

24. Thanks