This document summarizes Prof. Roshan Perera's presentation on emerging biotechnologies for developing antibodies, vaccines, and biosimilar products. It discusses active and passive immunity, types of vaccines and adjuvants, challenges in modern vaccinology. It also outlines a study showing that immunizing mice with a protein containing p-nitrophenylalanine generates neutralizing antibodies against wild-type TNF, protecting mice from LPS-induced lethality. This demonstrates a method for overcoming immune tolerance by incorporating unnatural amino acids. The presentation argues for establishing GMP facilities in Sri Lanka to produce monoclonal antibodies, vaccines, and conduct clinical trials to develop affordable treatments.

1. PROF. ROSHAN PERERA
KDU-CARE
INSTITUTE FOR COMBINATORIAL ADVANCED RESEARCH & EDUCATION
GENERAL SIR JOHN KOTELAWALA DEFENCE UNIVERSITY
Emerging Biotechnologies for the
Development and Manufacture of
Antibodies, Vaccines and Biosimilar
Products

2. Active and passive immunity
Active immunity: long-lasting protection (memory),
multiple effector mechanisms activated, lag time
Passive immunity: rapid protection, short duration
Abbas, Lichtman and Pillai. Cellular and Molecular Immunology, 7th edition, 2011

3. Types of Vaccines & Adjuvants
Vaccines
– Inactivated
– Live attenuated
– Chimeric
– Toxoid
– Subunit
– recombinant
– Conjugate
– DNA
Adjuvants
Inorganic compounds: alum, aluminum
hydroxide, aluminum phosphate, calcium
phosphate hydroxide
Mineral oil: paraffin oil
Bacterial products: killed bacteria Bordetella
pertussis, Mycobacterium bovis, toxoids
Nonbacterial organics: squalene, thimerosal
Delivery systems: detergents (Quil A)
Cytokines: IL-1, IL-2, IL-12

4. Challenges in modern vaccinology
• Polymorphism of antigens
• Variable immune responses
• Antigenic competition/interference in multivalent constructs
• Inability to enhance the immunogenicity of highly purified or recombinant
antigens
• Reduce the amount of antigen or the number of immunizations needed for
protective immunity
• Improve the efficacy of vaccines in newborns, the elderly or immuno-
compromised persons
• A major challenge in modern vaccinology is the development of robust methods
to selectively induce a strong immune response against self-proteins, or to
increase the immunogenicity of specific epitopes in foreign antigens that can
elicit neutralizing antibodies, but which are not immunodominant.
The goal of vaccination is to provide long-term protection against infection
through the generation of a strong immune response to the administered
antigen

5. Vaccines: the Phase III efficacy trials
• Malaria: RTS,S by GSK/US Army - efficacy 49%
• HIV: RV144, tested the “prime-boost” combination of
two vaccines: ALVAC® HIV vaccine (the prime) and
AIDSVAX® B/E vaccine (the boost). US Military -
efficacy 31.2%
• Dengue: CYD-TDV by Sanofi Pasteur - efficacy 56.5%
Strategic Goal: To develop 80% or more efficacious vaccine and
antibody candidates
INPIRATION

6. Natural & Unnatural Amino Acids
Genetically Incorporated Unnatural Amino AcidsNatural Amino Acids
IDEATION

7. Immunochemical termination of self-tolerance
A view of mTNF-α trimer
pNO2Phe
Serum titers for C57BL/6 mice immunization
—TNF is an important mediator in
inflammation, immune responses (i.e.
rheumatoid arthritis) and infection-related
phenomena
—but prolonged overexpression leads to
severe toxicity, which limits the use of TNF as
an anticancer agent
INVENTION

8. • Immunization (5 µg antigen/mouse)
• 23 C57BL/6 mice (age: 15 weeks)
• LPS dose 8.5 mg/kg on day 1
0 1 2 3 4 5 6 7 8
0
20
40
60
80
100
WT_mTNF
WT_mod_mTNF
PBS
days
Survival(%)
mTNF-Y86NO2F (n=8)
mTNF-wt (n=8)
PBS (n=7)
Lipopolysaccharide (LPS) challenge
—This immunization efficiently protects mice against a lipopolysaccharide (LPS) induced
lethality.
—Due to the close structure similarity, the elicited antibodies can cross-react with the
corresponding self-protein.
~90% survival rate
~10% survival rate
—The septic shock induced by LPS is known to involve in the production and release of TNF
INVENTION

9. Summary
• —Mice immunized with this pNO2Phe containing protein generate a strongly
neutralizing antibody response that effectively cross-reacts with wild-type
mTNF,
• —immunization efficiently protects mice against a lipopolysaccharide (LPS)
induced lethality.
• —These results strongly suggest that self-proteins bearing a unique NO2
group, a highly immunogenic moiety not found in natural occurring proteins,
will be recognized as foreign antigens by the immune system.
• —This approach should provide a general method for breaking immune
tolerance of self-proteins and the productions of vaccines.
H2N CO2H
NO2
p-nitrophenylalanine (pNO2Phe)
INVENTION

10. National Immunization Programme
• The estimated cost of vaccines procured by
the government of Sri Lanka is over Rs. 400
million/year.
• Every year the cost of vaccine is increasing
substantially.
• Antibody treatment is also an expensive affair
(i.e. Rs 70,000/injection for Rheumatoid
arthritis)

11. Development of Vaccine Candidates, Monoclonal
Antibodies and Protein Therapeutics in Sri Lanka
A GMP certified vaccine and antibody production plant
with recombinant technology should be set up in Sri
Lanka for:
1. Production of high quality disease specific monoclonal
antibodies and virology (infectious) reagents for
research and diagnostics.
2. Production of antibodies for snake venom and
rheumatoid arthritis.
3. Development of vaccine candidates for rabies
4. Production of Biosimilar products

12. Preclinical & Clinical Trials
KDU-CARE and KDU University Hospital ($222 million project)
IMPLEMENTATION