Strategies for the Generation of Influenza Vaccine

VACCINES FOR DISEASES

WITH PANDEMIC POTENTIAL
Basic Influenza Virus Structure
Three categories of flu viruses : A, B or C depending
on the virus’s nucleoprotein.

Birds, pigs, etc. Humans only

Human, mild
symptoms
Influenza Virus A
• Can be further identified by various protein
spikes that dot the virus’s outer covering.
• 18 different hemagglutinin (H proteins) and 11
different neuraminidase (N proteins).
• H antigens govern the ability of the virus to bind
to and enter cells, where multiplication of the
virus then occurs (i.e. attaches to cell receptor
and initiates entry into cell).
• N antigens govern the release of newly formed
viruses from the cells.
• Named using an “H” plus a number, such as type H1,
H2, etc & N” plus a number, such as type N1, N2, etc.
eg H1N5, H2N3 etc.
The Influenza A Pandemic Incidences

Source: WHO
Flu Viruses Change

Flu viruses have the capacity to change both

slowly, through small genetic changes that are
passed down to daughter generations, and quickly,
through a process called “reassortment” that mixes
larger genetic segments from several viral strains to
create a new virus.

Both processes are important to influenza’s

success as a disease-causing organism during flu
seasons and in pandemics.
 Antigenic Drift Vs Antigenic Shift

Antigenic Drift
• Minor change in Hemagglutinin or
Neuraminidase.
• Point mutations during replication, but subtype
remains the same.
• Continuous changes, so virus continues to
cause illness and deaths.
• Limited partial immunity may exist to the
changed virus.
• Antigenic drift results in the need to update the
seasonal flu vaccines annually.
 Antigenic Drift Vs Antigenic Shift

Antigenic Shift
• Major change in HA / NA
• New subtype introduced.
• Caused by genetic re-assortment when 2
subtypes infect a host simultaneously.
• OR
• Caused by direct transmission from birds or
other animals to humans
• New virus subtype, could potentially lead to an
epidemic or pandemic, since human do not have
any immunity against this newly emerging
strain.
Developing Vaccines for Influenza Viruses with Pandemic Potential

STRATEGIES FOR THE GENERATION

OF INFLUENZA VACCINE
 Egg-based Production of
Influenza Virus Vaccine (Seasonal Flu vaccine)
• Currently, influenza vaccine production (approx 90%) relies on
the use of embryonated chicken eggs to culture the virus.
• Although this process is well controlled and efficient, it has
some limitations:
• vaccine supply from the conventional egg-based
manufacturing system is limited
• strain identification to vaccine production takes time
• susceptible to a pandemic outbreak (bird flu) that could
threaten egg-substrate availability
• time consuming process (i.e from seeding egg with virus to
formulation would take approx 9 months)
Current Scenario in Influenza Vaccine Manufacturing

Licensed Pre-Licensure R&D

• Inactivated • Adjuvanted • Virus-Like

influenza inactivated Particles
vaccines vaccines* • DNA
• Whole • Recombinant Vaccines
virion HA vaccine • Universal Flu
• Split virion Vaccines
• Live-
attenuated
vaccines
 Pandemic Influenza Vaccine
Manufacturing Challenges
• Non-adjuvanted pandemic vaccines required
increased doses for a protective Hemagglutination
inhibition assay (HAI) response.
• Yield of pandemic vaccine production in eggs is
lower than seasonal strains.
• Manufacturers have to race against time once a
Pandemic episode is declared.
• Stockpiled vaccine could be of little value if the
virus mutates.
 VLP Platform for Producing Vaccine in the Event of an
Influenza Pandemic
• VLPs is another promising alternative to traditional influenza
vaccine formulations.
• Mimics the natural virus but lacks the viral genetic material.
• Easily recognized by the immune system, and are
considered to be safe.
• Production process is viral free and does not require any
high level containment facilities or complex inactivation
steps.
• Currently available influenza vaccines are either made of
inactivated virions or antigen fragments/soluble proteins
(subunit), influenza VLPs are particulates by nature and
display intact and biochemically active viral antigens on
their surface.
 Development of Influenza VLPs
• Enveloped influenza VLPs have been developed by several
laboratories or biopharmaceutical companies, and were
demonstrated to induce protective immunity during preclinical
and clinical studies. 
• Eg of Influenza VLPs composition: Hemagglutinin (HA),
Neuraminidase (NA), and Matrix (M1) Protein.
• Trivalent seasonal influenza VLP (H1N1 A/Brisbane/59/2007,
H3N2 A/Brisbane/10/2007, B/Florida/04/2006) by Novavax (MD,
USA)
• induced a robust hemagglutination inhibition (HAI) antibody
response against all three strains.
• functional antibody against the NA enzyme, as measured in the
sera of immunized subjects using a NA inhibition assay (NAI).
• **Inhibition of NA activity may be important in reducing the
spread and severity of influenza infection.
Influenza Vaccine Manufacturing in Insect Cells

Isolate RNA or synthesize

Identify Virus type/H & N
DNA

Infect cells with

recombinant baculovirus &
Clone target DNA
express VLPs that contain
fragments into baculovirus
Hemagglutinin (HA),
vector
Neuraminidase (NA), and
Matrix (M1) Protein

Proteins self-assemble as
particles that resemble
influenza virus, but do not
contain flu RNA
Advantage of Recombinant VLP as Influenza Vaccine
• Speed from strain selection to product release is
weeks.
• Exact genetic match.
• Recombinant VLP’s are clinically proven (HPV,
HBsAg) with a broad immune response.
• No eggs required.
• Yields are higher than egg-based production;
potential for additional increase in yield.
• No pathogenic virus in manufacturing.
• Controlled cell culture process (Serum-free, Protein-
free, Suspension Culture).