Evaluating Synthetic Biology Produced Triterpene Oils in Adjuvant Formulations

ABSTRACT
Vaccine adjuvants help stimulate the body to generate an immune response when co injected with an antigenthe compound for which immunity is desiredAlthough the human body is capable of producing antibodies to
most injected antigensadjuvants can help boost the body s ability to mount a successful and acquired immune
response to the presented targetAdjuvants have been shown to enhance immunity against targets that usually
exhibit poor immune responses such as highly purified and recombinantly produced antigensThis is
particularly important for individuals which may have a less robust immune systemsuch as children or the elderlyOne of the well studied adjuvants that is approved for use in humans nearly worldwide is the oil in water emulsionMFThe oil used in this formulation is the hydrocarbon triterpenesqualeneSqualene is a natural oil produced
in all eukaryotic organisms as it is the precursor to sterolse gcholesterolMFis a component of Fluadan
FDA approved seasonal influenza vaccine for peopleyears oldThe traditional source of squalene has been
from shark liver oilwhich is no longer viable nor sustainableIn light of these limitationsand the desire to
develop more efficacious adjuvantsfermentable yeast and transgenic plants have been engineered to produce
high amounts of squalene and several relatednovel triterpenesThese include botryococcene plus monoand
di methylated forms of squalene and botryococceneThe experiments proposed here are designed to
accomplish two specific aimsusing a mouse modeldemonstrate that our Synthetic Biology generated
squalene performs equivalent to shark derived squalene in adjuvant vaccine formulationsandevaluate ournovel triterpenes in adjuvant formulations for their abilities to induce elevated or altered immune responses
compared to squaleneOur studies will quantitatively compare the immunogenicity of a trivalent influenza
vaccine plus and minus the different triterpene adjuvant formulations by measuring antibodyIgGIgG a and
IgEand hemagglutination inhibitionHItitersThe possibility that our antigen triterpene adjuvant formulations
might induce undesirable effects will be assessed by measuring inflammatory cytokine levels in sera samples of
the inoculated animalsOnce the feasibility of using these compounds in an adjuvant system is demonstrated in
this Phase I studythe foundation for several follow on Phase II studies will be establishedonedemonstrating
the biological equivalency of our Synthetic Biology produced squalene would endorse efforts to use process
engineering methodologies to scale up our squalene manufacturing capabilities for marketplace
commercializationand twoif any of our novel triterpenes proves efficacious in stimulating antibody titerswe
would need to pursue additional pre clinical evaluations of their safety and potential for inciting undesirable
inflammatory responses PROJECT NARRATIVE
Vaccine adjuvants play a special role in the development of adaptive immunological responses because they
stimulate more robust titers of antibodies without inciting undue inflammationAlthough triterpene based
adjuvants are recognized for their efficacy in stimulating desirable immune responselimiting sources of
squalene and squalene analogs have impeded use and development of these oil in water adjuvantsWe will
provide equivalency testing of new Synthetic Biology generated triterpenespaving the way for scale up process
engineering towards commercialization and providing the justification for advanced pre clinicalsafety
evaluations