Animals in the Production of Vaccines and Pharmaceuticals – Hybridomas and Monoclonal antibodies
Introduction
Animals have played a crucial role in the production of vaccines and pharmaceutical products. They serve as biological systems for the synthesis of complex proteins, antibodies, hormones, enzymes, and vaccines. Animal-based production is especially important when post-translational modifications are required. Hybridoma technology and monoclonal antibodies represent major breakthroughs in medical biotechnology.
Animals in the Production of Vaccines
Role of Animals
Animals are used:
As hosts for propagation of pathogens
For testing vaccine safety and efficacy
For large-scale production of antigens
Types of Vaccines Produced Using Animals
Live attenuated vaccines
Example: Rabies vaccine (grown in chick embryo)
Inactivated (killed) vaccines
Example: Polio vaccine
Toxoid vaccines
Example: Tetanus, Diphtheria
Subunit vaccines
Antigenic parts isolated from pathogens
Common Animal Systems Used
Chick embryo
Mice
Rabbits
Horses
Monkeys
Examples
Rabies vaccine – chick embryo cell culture
Influenza vaccine – embryonated chicken eggs
Antisera – produced in horses
Animals in the Production of Pharmaceuticals
Animals are used to produce:
Hormones
Enzymes
Blood products
Antibodies
Examples
Product
Animal Source
Insulin (earlier)
Pig, Cow
Growth hormone
Animal pituitary
Antivenom
Horse
Blood clotting factors
Humanized animals
Antibodies
Mouse, Rabbit
Hybridoma Technology
Definition
Hybridoma technology is a method used to produce monoclonal antibodies by fusing antibody-producing B-lymphocytes with immortal myeloma cells.
Developed By
Georges Köhler and César Milstein (1975)
Nobel Prize – 1984
Steps in Hybridoma Technology
1. Immunization of Animal
Mouse is injected with a specific antigen.
B-cells in spleen produce antibodies.
2. Isolation of Spleen Cells
Antibody-producing B-lymphocytes are collected.
3. Fusion with Myeloma Cells
B-cells fused with cancerous myeloma cells using polyethylene glycol (PEG).
Forms hybridomas.
4. Selection of Hybridomas
Grown in HAT medium.
Only hybrid cells survive.
5. Screening
Hybridomas producing desired antibody are identified.
6. Cloning
Selected hybridomas cloned to obtain identical cells.
7. Large-scale Production
Cultured in bioreactors or injected into mice (ascites method).
Monoclonal Antibodies (mAbs)
Definition
Monoclonal antibodies are identical antibodies produced by a single clone of hybridoma cells and are specific to a single antigenic determinant (epitope).
Characteristics of Monoclonal Antibodies
Highly specific
Uniform in structure
Reproducible
Large-scale production possible
Applications of Monoclonal Antibodies
1. Medical Applications
Cancer diagnosis and therapy
Autoimmune diseases
Organ transplant rejection control
Targeted drug delivery
2. Diagnostic Uses
ELISA
Pregnancy test kits
Detection of pathogens
3. Research Applications
Identification of cell surface markers
Protein purification
4. Pharmaceutical Uses
Therapeutic antibodies (e.g., Rituximab, Trastuzumab)
Advantages of Animal-based Production
Proper protein folding
Post-translational modification
High biological activity
Limitations
Ethical concerns
Risk of contamination
High production cost
Immune reactions (HAMA response)
Conclusion
Animals play an indispensable role in the production of vaccines and pharmaceuticals. Hybridoma technology revolutionized antibody production, leading to the development of monoclonal antibodies with wide applications in medicine, diagnostics, and research. Despite limitations, advances in biotechnology continue to improve safety and efficiency.
50 MCQ
1. Hybridoma technology was developed by
a) Watson and Crick
b) Paul Ehrlich
c) Köhler and Milstein
d) Jenner
✅ Answer: c
2. Hybridoma cells are formed by fusion of
a) T cells and B cells
b) B cells and myeloma cells
c) Myeloma and stem cells
d) Plasma cells and T cells
✅ Answer: b
3. Myeloma cells are
a) Normal B cells
b) Cancerous antibody-producing cells
c) Cancerous plasma cells
d) Stem cells
✅ Answer: c
4. PEG is used in hybridoma technology for
a) Cell culture
b) Screening
c) Cell fusion
d) Cloning
✅ Answer: c
5. HAT medium is used for
a) Fusion
b) Selection of hybridomas
c) Antibody purification
d) Immunization
✅ Answer: b
6. Monoclonal antibodies are
a) Different antibodies
b) Polyclonal antibodies
c) Identical antibodies
d) Mixed antibodies
✅ Answer: c
7. Source of B-lymphocytes in hybridoma technology
a) Liver
b) Bone marrow
c) Spleen
d) Thymus
✅ Answer: c
8. Animal commonly used in hybridoma technology
a) Rabbit
b) Mouse
c) Horse
d) Monkey
✅ Answer: b
9. Antigen is injected into which animal
a) Goat
b) Rat
c) Mouse
d) Horse
✅ Answer: c
10. Ascites method is used for
a) Vaccine production
b) Antibody purification
c) Large-scale mAb production
d) Screening
✅ Answer: c
11. Monoclonal antibodies recognize
a) Multiple epitopes
b) Single epitope
c) Whole antigen
d) Pathogen
✅ Answer: b
12. Polyclonal antibodies are produced by
a) Single B cell clone
b) Multiple B cell clones
c) Hybridoma cells
d) Myeloma cells
✅ Answer: b
13. ELISA uses
a) Enzymes
b) Antibodies
c) Antigens
d) All of the above
✅ Answer: d
14. Antivenom is produced in
a) Cow
b) Horse
c) Sheep
d) Mouse
✅ Answer: b
15. Influenza vaccine is produced using
a) Mouse cells
b) Bacterial cells
c) Chick embryo
d) Horse serum
✅ Answer: c
16. Rabies vaccine is produced in
a) Monkey kidney cells
b) Chick embryo
c) Human liver cells
d) Yeast cells
✅ Answer: b
17. Insulin was earlier extracted from
a) Sheep
b) Cow and pig
c) Mouse
d) Horse
✅ Answer: b
18. Toxoid vaccine example
a) Polio
b) Rabies
c) Tetanus
d) Measles
✅ Answer: c
19. Hybridomas are
a) Immortal cells
b) Short-lived cells
c) Dead cells
d) Stem cells
✅ Answer: a
20. Main advantage of monoclonal antibodies
a) Low specificity
b) High specificity
c) Low cost
d) Mixed response
✅ Answer: b
21. Myeloma cells lack ability to
a) Divide
b) Produce antibodies
c) Survive in culture
d) Fuse
✅ Answer: b
22. Nobel Prize for hybridoma technology was awarded in
a) 1975
b) 1980
c) 1984
d) 1990
✅ Answer: c
23. Pregnancy test kit works on
a) Antigen-antibody reaction
b) DNA hybridization
c) PCR
d) Cell fusion
✅ Answer: a
24. Monoclonal antibodies are used in
a) Cancer therapy
b) Diagnosis
c) Research
d) All of the above
✅ Answer: d
25. HAMA reaction occurs due to
a) Mouse antibodies in humans
b) Human antibodies in mice
c) Viral infection
d) Bacterial contamination
✅ Answer: a
26. Antibody-producing cells are
a) T lymphocytes
b) B lymphocytes
c) Macrophages
d) Neutrophils
✅ Answer: b
27. Which vaccine uses killed pathogen
a) Live vaccine
b) Inactivated vaccine
c) Subunit vaccine
d) Toxoid
✅ Answer: b
28. Monoclonal antibodies are uniform because they are produced from
a) Different cells
b) Same clone
c) Different antigens
d) Multiple species
✅ Answer: b
29. Cell fusion in hybridoma is chemical mediated by
a) Ethanol
b) PEG
c) Methanol
d) Acetone
✅ Answer: b
30. Animal used for antiserum production
a) Mouse
b) Horse
c) Rabbit
d) Monkey
✅ Answer: b
31. Diagnostic use of mAbs includes
a) Blood grouping
b) ELISA
c) Cancer markers
d) All
✅ Answer: d
32. Vaccine containing weakened pathogen
a) Killed
b) Toxoid
c) Live attenuated
d) Subunit
✅ Answer: c
33. Main limitation of animal-based pharmaceuticals
a) Ethical issues
b) High specificity
c) Easy availability
d) Low cost
✅ Answer: a
34. Hybridoma cells can be cultured
a) In vitro
b) In vivo
c) Both a and b
d) None
✅ Answer: c
35. Blood clotting factor VIII is used in
a) Diabetes
b) Hemophilia
c) Cancer
d) Malaria
✅ Answer: b
36. Subunit vaccines contain
a) Whole pathogen
b) Toxin
c) Antigenic part
d) DNA
✅ Answer: c
37. Primary host used in vaccine safety testing
a) Human
b) Animal
c) Plant
d) Microbe
✅ Answer: b
38. Hybridoma technology produces
a) Polyclonal antibodies
b) Monoclonal antibodies
c) Antigens
d) Vaccines
✅ Answer: b
39. Function of antibodies
a) Digestion
b) Immunity
c) Respiration
d) Excretion
✅ Answer: b
40. Which is NOT an application of monoclonal antibodies
a) Cancer therapy
b) Hormone production
c) Diagnosis
d) Research
✅ Answer: b
41. Antigen specificity of monoclonal antibodies is
a) Low
b) Moderate
c) High
d) Variable
✅ Answer: c
42. Ascitic fluid is rich in
a) Antigens
b) Antibodies
c) Hormones
d) Enzymes
✅ Answer: b
43. Vaccine against tetanus is
a) Live
b) Killed
c) Toxoid
d) Subunit
✅ Answer: c
44. Which animal is commonly used for polyclonal antibodies
a) Mouse
b) Rabbit
c) Horse
d) All
✅ Answer: d
45. Hybridoma cells are selected by
a) PEG
b) HAT medium
c) Antigen
d) Enzyme
✅ Answer: b
46. Monoclonal antibodies are used to identify
a) DNA
b) RNA
c) Cell surface markers
d) Lipids
✅ Answer: c
47. Production of vaccines requires
a) Animals
b) Microorganisms
c) Cell culture
d) All of the above
✅ Answer: d
48. First monoclonal antibody produced in
a) Human
b) Mouse
c) Rabbit
d) Horse
✅ Answer: b
49. Myeloma cells are
a) Mortal
b) Immortal
c) Dead
d) Differentiated
✅ Answer: b
50. Monoclonal antibodies are preferred because they are
a) Cheap
b) Non-specific
c) Highly specific
d) Unstable
✅ Answer: c
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