I am a pharmacist. These slides describe biotechnology topic. I hope students get good benefits about it.

2.  It is the use of biological organisms, systems, or
processes to develop technologies and products to
improve the quality of life.
 Biotechnology is used in various fields including
agriculture, food science, and pharmaceuticals.
 Pharmaceutical Biotechnology companies use
recombinant DNA technology, which includes genetic
modification of cells, or a monoclonal antibody for
making their biotechnological products.

3.  Biotech is the term used for biotechnology or products
produced by biotechnology. These drugs are also called
biological, biotech drugs, biological drugs, or
biopharmaceuticals.
 True biotech products are manufactured in live biological
systems known as expression systems.
 Examples of Classes of Protein-based Biotech drugs,
Erythropoietin (EPO)
Blood factors (Factor VII)
Human Growth Hormone: HGH, Somatotropin,
Cytokines
i) Interleukins
ii) Interferon

4.  Medical Biotechnology is the use of living cells and
cell materials to research and produce pharmaceutical
and diagnostic products that help treat and prevent
human diseases.

5.  Humilin: It is form of insulin produced by genetic
engineering of insulin gene into E.Coli. First marketed
by Eli-Lilly in 1982. Humilin structure is similar to
human insulin but it’s alpha and beta chains are
synthesized separately in different strains of E.Coli
 Interferons ( Pegasis, Uniferon, Roferon): It is
used in leukemia and kaposis sarcoma and is now
produced by DNA Recombinant technology using
E.Coli as host.
 Human Growth Hormones: A human growth
hormone (Protropin) is also produced genetic
engineering which is used to treat pituitary dwarfism.

6.  Anti Hemophilic Factor: Factor VIII whose deficiency
causes hemophilia A is now available for use in patient
with hemophilia by using this technology.
 tPA protein: A thrombolytic agent called as tissue
plaminogan activator (tPA) is prepared for use as drug to
dissolve blood clot which are the cause of heart attacks
and strokes.
 Vaccines: Genetically engineered vaccines for many
infectious diseases. Most common is recombinant
hepatitis B vaccine which contains a surface protein
from the hepatitis virus. This protein is synthesized by
recombinant yeast cells and then purified by injection.
The HBV vaccine is the much safer to use than a
weakened form of the actual virus.

7.  Interleukin-2: It activates immune system, used in treating
cancer with immune disorder.
 Tumor necrosis factor: It is also produced by genetic
engineering to treat tumor.
 Erythropoietin: It is glycoprotein hormone that plays role in
erythropoiesis and RBC production so recombinant
erythropoietin is used to treat anemia.
 Beta-Endorphin: it is a natural opiate used for pain relief is
also prepared genetically.
 Agriculture field: In agriculture, it is used to develop pest,
drought resistance plants and to increase crop production and
its nutrient value.
 Dairy Industry: Bacteria produced bovine growth hormone for
increase in milk production in cow, Recombinant rennin used
for manufacturing of cheese.
 Medical Laboratory: Gene probes are used for diagnostic
purposes of genetic disorders and infectious agents like HIV.

8.  Pharmacist play their role as knowledgeable
professional for the distribution, storage and
administration of the biotechnological products.
 Majority of biological products are proteins and
cannot be administered orally. Therefore pharmacist
should be aware of route of administration for the
particular biological product, furthermore in case
of powdered dosage form reconstitution of these
products require aseptic techniques.
 Because of unique qualities of these products storage
requirements will be likely to be specialized such as
large refrigeration areas or separate storage sections
regularly monitored by pharmacist

9.  Patient counseling is another factor in which
pharmacists are actively invovled. Owing to unique
and complex properties of biotech products, patients
must receive in depth information about : dilution,
reconstitution, administration, storage and
adverse effects.
 Pharmacists are also involved in clinical monitoring
of the biological products. A potential role will be
found in ambulatory clinics where administration of
long term monitoring of these products will take
place.

10.  Owing to high cost of the biological products,
pharmacist must become involved in administrative
aspects of drug use process.
 There must be a formulary system that is capable of
handling requests for these agents. It is duty of
pharmacist to ensure rationale safe and cost effective
therapy is provided to all patients.
 Research and development of new biotechnological
products require specialized training and view point of
pharmacist.
 Pharmacist are also involved in ethics and safety issues
of the new product monitoring it for all the safeguards
and to avoid genetically engineered organisms into
environment or exposing human to any risk of
infection.

11. 1. Isolation of gene of interest.
2. Introduction of gene to expression vector.
3. Transformation into host cells.
4. Growth of cells
5. Isolation & purification of protein.
6. Formulation of protein product.

12.  Genes are pieces of DNA which store information for making
specific proteins that control specific traits. Genes are present
in nucleus chromosomes which code for one polypeptide.
 The double strand of DNA are cut open at specific site i.e
Nucleotide sequence by special enzymes which are known as
Restriction Endonucleases.
 Restriction enzymes are molecular scissors that cut DNA into
pieces. Find out which enzymes will cut, and where by making
a restriction map.
 Ligases are the enzymes used to joined pieces of DNA through
covalent bonds.
 Through these enzymes DNA containing genes of interest is
cut and joined with vector.
 Vector (molecular biology) In molecular cloning,
a vector is a DNA molecule used as a vehicle to artificially
carry foreign genetic material into another cell, where it can be
replicated and/or expressed (e.g.- plasmid, cosmic, Lambda
phages). A vector containing foreign DNA is termed
recombinant DNA.

14.  DNA recombination is a process in which the pieces of DNA
from different organisms are artificially mixed to create
Recombinant DNA.
 Steps involved in Recombinant DNA technology are
1. DNA extraction
2. Purification
3. Fragmentation.
 Fragmentation of DNA is done by specific 'restriction'
enzymes and is followed by sorting and isolation of
fragments containing a particular gene. This portion of the
DNA is then coupled to a carrier molecule called as
PLASMID (vector).
 The hybrid DNA is then introduced into a chosen cell for
reproduction and synthesis.

16. Different organism may be used to express a target protein, the
expression vector used therefore will have elements specific for
use in the particular organism.
An expression vector, otherwise known as an expression
construct, is usually a plasmid or virus designed for gene
expression in cells. The vector is used to introduce a specific
gene into a target cell, and can commander the cell's mechanism
for protein synthesis to produce the protein encoded by the gene.
The most commonly used organism for protein expression is the
bacterium, Escherichia coli. However not all proteins can be
successfully expressed in E. coli, and other systems may therefore
be used such as
 Yeasts e.g commonly used for protein expression is Pichia
pastoris.
 Baculovirus.
 Mammalian cell culture.

17.  After the transfer of rDNA into host of choice , i.e
bacteria, yeast, Ecoli they are cultivated in culture
medium.
 For research (small scale):
 Cell culture flasks
 For productions (large scale):
 Fermentors and Bioreactors

18.  It is important to provide nutritional
conditions that exist in bacterial natural
habitat.
 Common components:
• Water
• Source of carbon and energy
• Source of nitrogen
• Trace elements
• Growth factors
• Buffer
• For growth on small scale incubators are used ,the most
commonly used is shake flask incubator.

19.  Bioreactor or fermentor is a container in which substrate is
turned into product, where gene of interest is mass
produced.
 Production Process:
 Types of production processes are:
1. Batch
2. Continuous
3. Fed Batch
 The mode of feeding determines the classification
of the bioreactor.

20.  In this process the bioreactor is only fed once.
 The bioreactor will be allowed to run till completion
 Very difficult to achieve in real life because there
should be no input to or withdrawal from the
bioreactor even for sampling.

21. ADVANTAGES
 Ease in operating.
 genetic stability of organism could be controlled if it is
genetically engineered biocatalyst.
 lower contamination risk.
Disadvantages
 non-productive down time.
 batch to batch variability is problem.
 accumulation of inhibitory product.

22.  The bioreactor is fed continuously
 The amount of feed introduced into the bioreactor
equals the removed volume.
 The process is sensitive and subjected to influence
from various factors.

23. Advantages
 efficient, higher productivity.
 Uniform quality of product.
 no accumulation of inhibitory products.
Disadvantages
 destruction of biocatalyst.
 higher contamination risk.

24.  In fed Batch process , one or more nutrients are fed to
the bioreactor during cultivation and in which the
product remain in the bioreactor until the end of the
run.
 Possible to control the rate of growth of the
microorganisms or the concentration of the biomass
by controlling the feed parameters.
 most commonly used process in industry.

25.  There is necessity for the isolation and purification of
protein prepared for Biotech drugs from micro
organisms.
 The proteins required may be extracellular or
intracellular so for isolation of extracellular products
destruction of cell is not necessary but for intracellular
cell disruption is needed which is done through
following methods
 Detergents lysis
 Enzymatic lysis
 Osmotic lysis
 Freeze-thaw cycles
 Ultrasonication
 Homogenization

26.  Differential centrifugation(Separation of proteins or
any material on the basis of their size, mass, and
density)
 Dialysis (Based on osmotic pressure)
 Differential solvent precipitation
 Gel electrophoresis
 Column chromatography