1. Purified proteins Recombinant proteins Whole inactivated or attenuated organisms DNA encoded antigens Bacterial products Toxins Cytokines, peptides Oil in water emulsions Aluminum compounds Mineral salts Polymers Antigens Immune Potentiators Delivery Carriers Modern Vaccine Components

2. MOTIVATION A Modular approach to constructing vaccines Adaptor Biodegradable polymer Recognition elements Antigen Protective transport elements Endosomal Disruption element S S Protection during transport Dendritic Cell recognition Encapsulated antigen subunit Encapsulated Endosomal disruptor Transepithelial cell transport 100-400 nm

3. Biodegradable polymer nanoparticles Encapsulated antigen subunit 100-400 nm A Sustained release of encapsulated antigen from a biodegradable core. 50/50 Poly (lactide-co- glycolide) polymer

4. Adaptor Proteins 100-400 nm B High density surface presentation of adaptor elements

5. Adaptor Proteins Surface presentation is long-lived with sustained release of antigen Biotin-PE C Long-lived surface presentation

6. 100-500 nm Polysaccharide Backbone TLR4 Ligand Lipid A Anchor + Antigen-loaded Biodegradable Nanoparticle Antigen-Delivery to Dendritic Cells: Targeting Toll Like Receptor (TL4)

7. 100-500 nm Polysaccharide Backbone TLR4 Ligand Lipid A Anchor + Antigen-loaded Biodegradable Nanoparticle Antigen-Delivery to Dendritic Cells: Targeting Toll Like Receptor (TL4) LPS/OVA Uncoated/OVA Blank Inject SubQ Isolate Splenocytes Stimulate on an OVA coated Plate 7 days OR Oral Feeding

8. 100-500 nm Polysaccharide Backbone TLR4 Ligand Lipid A Anchor + Antigen-loaded Biodegradable Nanoparticle Antigen-Delivery to Dendritic Cells: (Subcutaneous Route) Targeting Toll Like Receptor (TL4) Stimulation of Splenocytes from vaccinated mice with OVA antigen 0.2 0.25 0.3 0.35 0.4 0.45 0.5 050100150 OVA (ug/ml) Blank Particles Unmodified Particles/OVA LPS Particles/OVA Proliferation Index (Post 3 Days)

9. 100-500 nm Polysaccharide Backbone TLR4 Ligand Lipid A Anchor + Antigen-loaded Biodegradable Nanoparticle Antigen-Delivery to Dendritic Cells: (Oral Route) Targeting Toll Like Receptor (TL4) Oral Vaccination followed by OVA antigen Stimulation of Splenocytes OVA (ug/ml) Blank Particles Unmodified Particles/OVA LPS Particles/OVA Proliferation Index (Post 3 Days) 0.3 0.4 0.5 0.6 0.7 0.8 0.9 0306090120150

10. pH < 5 37 C Elastin (Expanded State) Elastin-Coated nanoparticle 200-500 nm pH > 5 37 C pH = 7pH = 2 0 0.005 0.01 0.015 0.02 020406080100 Time (hrs) Drug released (mg) Elastin (pH=2) No Elastin (pH= 2) Surface modification Protection during Transport: Surface modification with pH responsive polypeptides:

11. PATHFORWARD Optimal Assembly of modular units Transport Recognition of DC subsets Antigen presentation Ira Mellman Mark Saltzman Michael Caplan Robert Samstein Gilbert Addo Stacey Demento Erin Steenblock Thanks to:

12. Adaptor Biodegradable polymer Recognition elements Antigen Endosomal Disruption element Dendritic Cell recognition Encapsulated antigen subunit Encapsulated Endosomal disruptor Transepithelial cell transport 100-400 nm D Mediating transport and recognition