1. Introduction 2. Therapeutic approaches 3. Diagnostic Approaches 4. Antibiotics 5. Antibodies 5. Biopharmaceuticals 6. CRISPR/Cas9 gene editing in humans 7. Organ on chip 8. Sequencing of next generation THANK YOU

1. THERAPEUTIC & DIAGNOSTIC
APPROACHES IN
BIOTECHNOLOGY
PRIYANSHA SINGH (PC2021-14226)
Biotechnology in Pharmaceutical Sciences (BT 510) Unit- 9
M.S. (Pharm)- PHARMACOLOGY & TOXICOLOGY- NIPER GUWAHATI

2. Biotechnology in pharmaceutical
industry
 Diagnostics
 Antibiotics
 Vaccines
 Antibodies
 Biopharmaceuticals (insulin, interferon, GSF, CSF, therapeutic proteins etc.)
 Commercial aspects

3. INTODUCTION TO THERAPEUTIC APPROACHES
OF BIOTECHNOLOGY
 Medical Biotechnology is the use of living cells and cell materials (DNA) to
research and produce pharmaceutical/therapeutic products that help treat
and prevent human diseases.
 Biotechnology based drugs are being used in cancer, HIV/AIDS and other Auto-
Immune disorders.
 Biopharmaceutical drugs are referred to those drugs generated through
researches in cell biology, genetics and rDNA technology.
 rDNA and MAb are great opportunities for drug development and as new
approaches in drug delivery.

4. MAJOR AREAS OF THERAPEUTIC
APPROACHES OF BIOTECHNOLOGY
 GENE THERAPY
 NEW GENE THERAPY APPROACHES
 VACCINES
 STEM CELLS & THERAPEUTIC CLONING
 TISSUE ENGINEERING & XENOTRANSPORTATION
 DRUG DELIVERY & NANOTECHNOLOGY
 BIOPHARMACEUTICALS (eg.- INSULIN, GROWTH HORMONE, FACTOR VIII)

5. 1. GENE THERAPY/ GENE PILL
 Gene therapy is a collection of methods that allows correction of a gene defect that
has been diagnosed in a child/embryo. Here genes are inserted into a person’s cells
and tissues to treat a disease.
 Correction of a genetic defect involves delivery of a normal gene into the individual or
embryo to take over the function of and compensate for the non-functional gene.

6. Commercially available Gene Therapy
Products

7. 2. VACCINES
 Vaccines contain weakened or inactive parts of a particular organism (antigen) that triggers an
immune response within the body. Newer vaccines contain the blueprint for producing antigens
rather than the antigen itself. Regardless of whether the vaccine is made up of the antigen itself
or the blueprint so that the body will produce the antigen, this weakened version will not cause
the disease in the person receiving the vaccine, but it will prompt their immune system to
respond much as it would have on its first reaction to the actual pathogen.
 COMMERCIALLY AVAILABLE VACCINES

8. BIOPHARMACEUTICALS
 Biopharmaceuticals are complex medicines made from living cells or
organisms, often produced using cutting-edge biotechnological methods.

9. 3. ERYTHROPOIETIN
 Production of human clotting factors was
enhanced through recombinant DNA
technology.
 Human clotting factor IX was the first to be
produced through recombinant DNA
technology using transgenic Chinese hamster
ovary cells in 1986.
 Plasmids containing the factor IX gene, along
with plasmids with a gene that codes for
resistance to methotrexate, were inserted
into Chinese hamster ovary cells via
transfection
 COMMERCIALLY AVAILABLE FACTOR VIII:
NUWIQ 250 IU RANGE, NovoEight (Novo
Nordisk)

10. 4. INSULIN
 Production of genetically engineered human
insulin was one of the first breakthroughs of
biotechnology in the pharmaceutical industry.
 Insulin was first produced in Escherichia coli
through recombinant DNA technology in 1978.
 PRINCIPLE:- Mass production of human proteins,
vaccines, etc. by genetically modifying bacteria
or viruses.
 PROCESS: The human gene for insulin is placed
into bacteria, are cultured and allowed to
produce insulin which is collected, purified and
sold to diabetics worldwide.
 COMMERCIALLY AVAILABLE INSULIN: HUMALOG
(ELI LILLY)

11. 5. INTERFERON
 Interferons (IFNs) 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.

12. Marketed Interferons

13. 6. HUMAN GROWTH HORMONE
 Production of human growth hormone was first done in
1979 using recombinant DNA technology.
 scientists produced human growth hormone by
inserting DNA coding for human growth hormone into a
plasmid that was implanted in Escherichia coli
bacteria.
 This gene that was inserted into the plasmid was
created by reverse transcription of the m RNA found in
pituitary glands to complementary DNA.
 Prior to this development, human growth hormone was
extracted from the pituitary glands of cadavers, as
animal growth hormones have no therapeutic value in
humans.
 COMMERCIALLY AVAILABLE HGH: SOMATROPIN
(Novartis)

14. 7. MONOCLONAL ANTIBODIES (MAb’s)
 Tumor cells that are able to replicate endlessly are fused with mammalian
cells that produce a specific antibody which result in fusion called hybridoma
that continuously produce antibodies.
 Those antibodies are named monoclonal because they come from only 1 type
of cell, which is the hybridoma cell.
 COMMERCIALLY AVAILABLE MAb’s:

15. 8. DNase 1
 Recombinant DNase I (RNase-free) is an endonuclease that catalyzes, to the
same degree, the random degradation of both single- and double-stranded
DNA and produces 5'-P terminal oligonucleotides. This enzyme does not
exhibit protease activity.
 COMMERCIALLY AVAILABLE DNase I: DNase I recombinant (ROCHE)

16. 9. TISSUE PLASMINOGEN ACTIVATOR (TPA)
 TPA is used to treat coronary thrombosis (thrombolytic agent)- Protease that attaches
to blood clots & induces other blood components to break down clot.- plasminogen
gets converted to plasmin which stimulates degradation of fibrin.
 Streptokinase once used for this purpose is derived from Streptococcal bacteria and it
must be delivered I.V. only. Urokinase can be an alternative but it has a risk of
hemorrhage.
 TPA does not compromise on blood clotting elsewhere therefore reduces the risk
internal hemorrhage
 Transported to the site of action via circulation.
 Produced in mammalian cell culture (not E.coli)

17. 10. ERYTHROPOEITIN
 It is a glycoprotein hormone that controls
erythropoiesis or RBC production.
 rDNA technology was used to insert a foetal liver
genomic library apaI fragment encoding for
human erythropoietin (epo) into bowes
melanoma cells.
 The cells expressed the erythropoietin gene was
secreted into the culture medium together with
the normally secreted tissue plasminogen
activator
 COMMERCIALLY AVAILABLE ERYTHROPOEITIN:
EPOGEN (AMGEN)

18. INTRODUCTION TO DIAGNOSTIC APPROACHES
Medical diagnosis is another application of biotechnology in the health sector. Many times the pathogen
concentration increases by the time the disease is diagnosed. Hence, early diagnosis and knowledge of
pathophysiology are essential for an effective cure. This can be achieved with the help of techniques
such as Recombinant DNA Technology, Polymerase Chain Reaction (PCR) and Enzyme-Linked Immuno-
sorbent Assay (ELISA), etc. Following are a few examples of some popular diagnostic kits in market.
The Indian in vitro diagnostic market was valued at $1240 Million in 2019 and is projected to reach $2060
Million by 2027 registering a CAGR od 7.5% from 2020 to 2027

19. 1. Blood Glucose Level Biosensors
 When blood or a glucose solution is placed in the strip, a chemical reaction occurs
inside it, generating a small electrical current proportional to the glucose
concentration. This current is constantly monitored while the strip is in place,
allowing the device to monitor when blood is placed.
 COMMERCIALLY AVAILABLE GLUCOMETER: ACCU-CHECK Active blood glucose
detection kit (ROCHE Pharma.)

20. 2. Malaria Detection Kit
 Immuno-chromatographic tests are based on the capture of parasite antigen
from peripheral blood using monoclonal antibodies prepared against a malaria
antigen target and conjugated to either a liposome containing selenium dye
or gold particles in a mobile phase.
 COMMERCIALLY AVAILABLE MALARIA DETECTION KIT: SD BIOSENSOR Malaria
detection kit

21. 3. Latex Kits
 One of its type i.e. CRP Latex kits is used for the measurement of C- reactive
proteins in a patient’s serum. When a person is injured or ill, the amount of
C- reactive proteins increase in the plasma & this is then a measure for
susceptibility to injury or illness.
 The C-Reactive Protein test is based on the principle of the latex
agglutination. When latex particles complexed human anti-CRP are mixed
with a patient's serum containing C reactive proteins, an visible agglutination
reaction will take place within 2 minutes
 COMMERCIALLY AVAILABLE LATEX CRP DETECTION KIT: CRP TEST KIT (Lorne
Laboratories UK)

22. 4. Pregnancy Detection kits
 Pregnancy tests work by detecting the presence of the hormone, human chorionic
gonadotropin (hCG). hCG is a hormone produced by cells in the placenta. Its
production starts from the point at which the developing embryo attaches to the
uterus, 6-12 days after conception.
 COMMERCIALLY AVAILABLE PREGANANCY DETECTION KITS: i- can (Piramal
Healthcare)

23. 5. COVID-19 Detection Kits
 One type of rapid diagnostic test (RDT) detects the presence of viral proteins (antigens) expressed by the COVID-19 virus in a
sample from the respiratory tract of a person. If the target antigen is present in sufficient concentrations in the sample, it will
bind to specific antibodies fixed to a paper strip enclosed in a plastic casing and generate a visually detectable signal, typically
within 30 minutes. The antigen(s) detected are expressed only when the virus is actively replicating; therefore, such tests are
best used to identify acute or early infection.
 How well the tests work depends on several factors, including the time from onset of illness, the concentration of virus in the
specimen, the quality of the specimen collected from a person and how it is processed, and the precise formulation of the
reagents in the test kits. Based on experience with antigen-based RDTs for other respiratory diseases such as influenza, in
which affected patients have comparable concentrations of influenza virus in respiratory samples as seen in COVID-19, the
sensitivity of these tests might be expected to vary from 34% to 80%.
 COOMERCIALLY AVAILABLE COVID DETECTION KIT: MYLAB’S COVISELF (ICMR approved)

24. 6. HIV Detection Kit
 HIV p24 antigens are manufactured and attached to the bottom of a plastic testing dish. The dish is then
washed with the blood sample. If antibodies (upside-down Y-shapes) for the p24 antigen are present then
they attach to the antigens in the dish. This gives an HIV positive result.
 COMMERCIALLY AVAILABLE HIV DETECTION KITS: HIV ELISA (Oscar Medicare Pvt. Ltd.)

25. 7. Syphilis Diagnostic Kit
 The Syphilis Test is a qualitative membrane device based immunoassay for the detection of
Treponema palladium antibodies (IgG and IgM) in whole blood, serum or plasma. In this test
procedure, recombinant Syphilis antigen is immobilized in the test line region of the device.
 COMMERCIALLY AVAILABLE PRODUCTS: Dipstick Test Syphilis/TP strip. (OSCAR MEDCARE Pvt. Ltd.)

26. 8. Febrile Antigen Kits
 The antigens are suspensions of killed bacteria, stained to enhance the reading of agglutination tests. The
blue stained antigens are specific to the somatic “O” antigens and the red stained antigens are specific to
the flagellar “H” antigens.
 Suspensions of Proteus OX2, OX19 and OXK are used to detect rickettsial antibodies.
 Rose Bengal is a slide agglutination test for the qualitative and semi-quantitative detection of anti-Brucella
antibodies in human and animal serum. The stained bacterial suspension agglutinates when mixed with
samples containing specific IgG oro IgM antibodies present in the patient sample.
 COMMERCIALLY AVAILABLE DETECTION KIT: FEBRILE DETECTION KIT (Lorne Labs, UK)

27. 9. Cholesterol detection Kit
 In the cholesterol assay protocol, cholesterol oxidase acts on free cholesterol to produce a chemical
which reacts with a probe to generate color (570 nm) and fluorescence (Ex/Em = 538/587 nm).
Cholesterol esterase is used to hydrolyze cholesteryl ester into free cholesterol.
 COMMERCIALLY AVAILABLE DETECTION KIT: Microsidd Quickcheck
 Cholesterol monitoring kit

28. 10. Hepatitis Detection Kit
 The HBV antibody test, sometimes called the anti-HBV test, looks for
antibodies to the hepatitis B virus in blood. Antibodies are chemicals released
into the bloodstream when someone gets infected.
 The test for hepatitis B uses a blood sample collected from a finger prick. The
kit contains: an alcohol swab to clean your finger. a small lancet to make a
tiny cut in your fingertip.
 COMMERCIALLY AVAILABLE PRODUCT: RAPIKIT HBsAg TESTING KIT (NecLife)

29. Diagnostic tools

30. Diagnostic tools

31. Next generation
sequencing

32. Sequencing used to detect cancer
Immune response genes are
operationally defined
as antigen-specific genes
that control the ability of an
animal to raise an immune
response, humoral or
cellular, to a particular
antigen. The antigen
specificity is a crucial aspect
of the definition.

33. Sources of Antibiotics

34. Antibiotics production

35. Journey of manufacturing of vaccine

36. COVID- 19 Vaccine

37. Recombinant DNA tech in Hepatitis B
vaccine

38. Vaccine production plant

39. Antibody production

40. BIOPHARMACEUTICALS-
Insulin, Interferon, GSF,
CSF & therapeutic
proteins

41. General technique of production of
biopharmaceutical

42. Human Insulin Production

43. Interferon Production by rDNA tech

44. CRISPR-
Cas9

45. CRISPR/Cas 9 gene editing in human cell

46. Ex-vivo v/s In-vivo CRISPR/Cas9

47. Organ on chip

48. Pharmacokinetics & Pharmacodynamics
on organ on chip

49. Setup of an Organ on chip

50. How an organ on chip is analogous to the
actual organ

51. Various roles played by organ on chip