Introduction

 Biotechnology: refers to application of biological

organisms, systems, or processes to manufacturing
and service industries.

 Useful for large scale production.

 Nearly 700 novel drugs from biotech.

 Generally: protien using rDNA or monoclonal AB.

Historical Perspective
 Mutations, Genetic Engineering: Physical and
Chemical Agents.
 1940: Antibiotics, Pure chemicals e.g.amino acids and
nucleotides.
 1973: rDNA and molecular cloning.
-Technology for transferring genes e.g. gene cloning, site
directed mutagenesis, rapid DNA sequencing,
hybridoma tech, monoclonal AB, protoplast fusion
and tissue culture.
Milestone
Double helix model for 3 D structure of
DNA
Cleavage of DNA by restriction
endonucleases
Determination of genetic code
Identification of DNA ligase
DNA cloning
Hybridoma creation
DNA sequencing Technology
US approval for Human Insulin derived
from DNA tech
Scientist
J, D. Watson and F.H. Crick 1952-53
W. Arber, 1962; M. Meselson and R.
yuan , 1968
M. Nirenberg, P. Heder and H. Khorana
1966
M. Gellert 1967
H. W. Boyer, S, Cohen and P. Berg 1971-
72
C. Milstein and G, Kohler
F. Sangar and W. Gilbert 1977
Genetech and Eli Lilly 1982
Terminology
 Gene cloning: technology for identifying, isolating and
copying a gene code for valuable gene

 -analysis, protein synthesis.

 -production of large quantities of any gene in any host.
 E.g. 1972 recombinant molecule formed by restriction
enzyme to fragments and subsequent ligation.
 -novel characteristics and large scale production of
vaccines, hormones, clotting factors.
 Site directed mutagenesis: point mutation at defines
loci. Possible only if nucleotide sequence is known.
Various agents.
 Rapid DNA Sequencing: gene characterization and
manipulation.
 -alter amino acid sequence.
 -SAR
 -modify physiological functions of protien.
 Hybridoma: Milstein and Kohler 1975.
 -Fusion of Myeloma and AB producing
lymphocytes to produce monoclonal AB.

 Efficiency of recombination increased with

protoplast fusion e.g. for synthesis of secondary
metabolites.
Scope and Application
 Prophylaxis, Therapy, Diagnosis and Discovery
 Viral Vaccines
 1. Bioactive molecules as Pharmaceuticals:
 -Hormone deficiency diseases:
 A) Insulin
 B) Human Growth Hormone
 C) Growth Releasing Factor
 D)Erythropoetin: kidney, hypoxia, red blood cell
precursors
 E) Blood clotting Factors VIII and IX.
 F) Interferons
 G)TNF and Lymphotoxin
 H)Bacterial streptokinase derived from human
urokinase and Tissue Plasminogen Activator can
dissolve blood clots.
 Bioactive peptides used as vaccines for hepatitis B,
malaria, herpes, influenza etc.
 2) Immunomodulators: improve responsiveness in
cancer
 -Identification of targets for designing: oncogenes and
receptors.
 Cloning of estrogen , insulin, muscarinic receptors.
 3) Monoclonal Antibodies: Hybridoma technology,
Diagnostic kits
 4)Hybrid AB: Immunotoxin by constructing single
chain AB toxin by fusion protein in E. coli. E.g
interlukin 2 receptor +ve cells in allograft rejection,
AIDS.
 5) Hybrid Antibiotics: same cell two biosynthetic
pathways. Structure different from parent.
 6) DNA probes: identification of particular sequence,
genes, fragments.
 -identification of pathogen and genetic disease.
 7) Gene Therapy: immune deficiencies, hereditiary
anaemias, urea cycle defects, metabolic defects,
deficiency of peptide hormones, cancer etc.
 8) Microbial Strain Improvement: Protoplast fusion-
hybrid anibiotics, desensitization of enzyme to
metabolic inhibition
 9) Biosensors: measurement of compounds in bio-
fluids. glucose electrodes.
 10) Plant cell culture: disease resistance, stress or
herbicides.
 N fixation from K. pneumonia.
 Pest resistance B. thuringinesis.
 11) Animal and Mammalian cell culture.
 -growth rate, milk yield, disease resistance:hormones
and monoclonal AB, Ca-DNA and viral vector,
fertilized egg.
 12)Enzymes: therapeutic, analytical, manipulative.
 -new altered genes and site directed mutagenesis.
 13) Drug delivery: Proteins broken down in body
 -Bio-erodible non toxic synthetic polymer 5-300µ.
 -Transdermal application Scopolamine via TDDS.
 -Liposomes:oral occular, injection.
 E.g. doxorubicin HCl, Rifampicin.