BIOTECHNOLOGY
It Is the technology that uses biological systems, living organisms or parts of this (organisms or enzymes)
to develop or create different products.
▪ Examples: wine and cheese-making, fermentation process that yields antibiotics, recombinant DNA
technologies
PHARMACOBIOTECHNOLOGY
This refers to the wide application of “biotechnology” to the development of pharmaceuticals or
pharmaceutically active substances.
▪ An example of pharmacobiotechnology product is recombinant lnsulin Ely Lilly and company and in
1982 Humulin was approved by the FDA.
FIELDS OF BIOTECHNOLOGY
1. Recombinant DNA–technology
▪ This technology permits the replication of a specific DNA sequence.
2. Molecular biology
▪ Chiefly concerns itself with understanding the interactions between the various systems of a cell,
including the interrelationship of DNA, RNA and protein synthesis and learning how these interactions
are regulated.
3. DNA-alteration
▪ It can be acquired/somatic or as genetic mutation
Mutations can also occur as the result of exposure to environmental factors such as smoking, sunlight
and radiation. Often cells can recognize any potentially mutation-causing damage and repair it before it
becomes a fixed mutation.
4. Gene-splicing
▪ Gene splicing is the process of chemically cutting DNA in order to add bases to the DNA strand.
5. Genetic engineering
▪ Genetic engineering is the process of using recombinant DNA (rDNA) technology to alter the genetic
makeup of an organism.
6. Immunology
▪ It is a branch of biology that covers the study of immune systems in all organisms.
7. Immunopharmacology
▪ A branch of pharmacology concerned with the application of immunological techniques and theory to
the study of the effects of drugs especially on the immune system
EXAMPLES OF BIOTECHNOLOGY-BASED PRODUCTS
Actimmune®
For chronic granulomatous disease
Betaseron®
For relapsing, remitting multiple sclerosis where the immune system attacks the protective sheath
(myelin) that covers nerve fibers
Epogen®
For treatment of anemia associated with chronic renal failure
KoGENate®
For treatment of haemophilia A. It is genetic deficiency in clotting factor VIII, which causes
increased bleeding and usually affects males
Leukine®
For autologus bone marrow transplantation (Autologous means obtained from the same
individual)
OncoScint® CR/OV
For detection, staging and follow up of colorectal and ovarian cancer
Proleukin®
For renal cell carcinoma.
IMPORTANT PROCESSES IN BIOTECHNOLOGY
1. Recombinant DNA
Steps:
1st Restriction enzymes cleave the DNA at particular sites
2nd enzymes known as ‘ligases’ will help to insert the cut piece of DNA (called the ‘insert’) directly into
the Vector DNA (i.e., viral DNA or plasmid).
3rd resulting vector DNA shall be capable of entering conveniently into a ‘host’ cell or otherwise called
as microorganism.
4th once the foreign DNA (or vector DNA) gains an entry into the microorganism (i.e., host organism), it
is known as a “recombinant organism”
2. Restriction Enzymes
▪ It is also known as ‘restriction endonucleases’.
▪ It acts as a pair of “molecular scissors and is found to ‘digest’ DNA into corresponding short strands at
very particular sites
3. DNA–Ligase
▪ It is a DNA-joining enzyme. If two pieces of DNA have matching ends, ligase can link them to form a
single, unbroken molecule of DNA.
▪ The DNA pieces cut that were with the same enzyme shall possess ‘sticky ends’ which would anneal
under the appropriate and favorable conditions when such ‘pieces’ are pooled together.
▪ Mixed pieces are treated with an enzyme, termed as DNA – ligase, it will ultimately give rise to an
intact piece of recombinant DNA by the combination of phosphodiester bond (at the ends of the DNA
strands) with the complementary bases (adenine, cytosine, guanine and thymidine).
4. Cloning Vector
▪ This is a small piece of DNA that can be stably maintained in an organism, and into which a foreign DNA
fragment can be inserted for cloning purposes.
▪ May be a plasmid or a bacteriophage
▪ Gene having a close resemblance to a particular protein shall be joined together with a cloning vector
so as to enable it get transferred into a host cell.
5. Hybridization Probes
▪ It refers to complementary sequence of DNA (either larger segments of DNA or synthetic
oligonucleotides or even whole plasmids) that is specifically labelled with anyone of the three different
means namely:
o Radioactive substance
o Fluorescent material
o Chromogenic material
6. Cloning Process
▪ Cloning simply means to make identical copies. It is the process of generating a genetically identical
copy of a cell or an organism. Cloning happens often in nature—for example, when a cell replicates itself
asexually without any genetic alteration or recombination.
HUMAN PROTEINS
A. Hormones
▪ Human Insulin
o Humulin® (Eli Lily)
o Recombinant DNA technology in E. coli (1983)
▪ Calcitonin
o thyroid hormone which helps in the regulation of calcium retention in bones
B. Blood Products
▪ Tissue Plasminogen Activator (tPA)
o It is aserine protease that catalyzes the conversion of plasminogen
to plasmin (plasmin is responsible for the breakdown of unnecessary and unwanted blood clots
▪ Factor VIII: The genetically engineered Factor–VIII
o Cloned in hamster cells and used for the treatment of hemophilia
▪ Lymphokines
a. α-Interferon –
b. Interleukin–2 –
c. Tumor Necrosis Factor (TNF) –
VACCINES
Vaccines are suspension of infectious agents, or some part of them given for the purpose of establishing
resistance to an infectious disease
Types of traditional vaccines
▪ containing living attenuated infectious organisms – example: vaccine for poliomyelitis
▪ containing soluble toxins of microorganisms (toxoids) –
▪ containing substances extracted from infectious agents –
Recombinant vaccines (or subunit vaccines)
▪ all the gene(s) solely dedicated for encoding the cell surface molecules that eventually trigger the
immune activation function is isolated
o these genes are removed from the pathogenic genome by endonuclease action and
subsequently inserted into a cloning vehicle that later on employed to transform either a
bacterium (E. coli) or yeast (Saccharomyces cerevisiae) host cell.
MONOCLONAL ANTIBODIES (MABs)
Hybridoma technology –
Antibody producing cells are duly fused with the respective tumor cells which results into the production
of a hybrid that could be grown in an appropriate laboratory culture and which reproduces antibodies of
a single specificity (monoclonal antibodies) with the longevity and reproductivity of the myeloma
DNA PROBES AND RELP ANALYSIS
DNA probes –
Restriction Fragment Length Polymorphism (RFLP) Analysis
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