A macromolecular delivery vehicle for protein-based vaccines: Acid-degradable protein-loaded microgels

Murthy, Niren; Xu, Mingcheng; Schuck, Stephany; Kunisawa, Jun; Shastri, Nilabh; Fréchet, Jean M. J.

doi:10.1073/pnas.0930644100

Cited by 389 publications

(376 citation statements)

References 23 publications

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“…19,22,38 As a model protein cargo, we examined the assembly of multilayers containing ovalbumin (ova), a 45 KDa globular protein routinely used as a model vaccine antigen. [39][40][41] We first prepared PEM films by alternating adsorption of Poly-1 and ova from aqueous buffers onto glass slides, utilizing electrostatic interactions between the protein and polymer to mediate assembly and monitoring protein incorporation via the absorbance of fluorophore-conjugated ova. Based on the pI of ova (~4.6) and the pK a of Poly-1 (between 4.5 -8), we tested film assembly at pH 5 -7, varying the pH of both Poly-1 and ova adsorption solutions ( Figure S2).…”

Section: Resultsmentioning

confidence: 99%

Layer-by-Layer-Assembled Multilayer Films for Transcutaneous Drug and Vaccine Delivery

Su¹,

Kim²,

Kim³

et al. 2009

ACS Nano

156

120

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We describe protein-and oligonucleotide-loaded layer-by-layer (LbL)-assembled multiplayer films incorporating a hydrolytically degradable polymer for transcutaneous drug or vaccine delivery. Films were constructed based on electrostatic interactions between a cationic poly(β-amino ester) (denoted Poly-1) with a model protein antigen, ovalbumin (ova), and/or immunostimulatory CpG (Cytosine-phosphate diester-Guanine rich) DNA oligonucleotide adjuvant molecules. Linear growth of nanoscale Poly-1/ova bilayers was observed. Dried ova protein-loaded films rapidly deconstructed when rehydrated in saline solutions, releasing ova as non-aggregated/non-degraded protein, suggesting that the structure of biomolecules integrated into these multilayer films are preserved during release. Using confocal fluorescence microscopy and an in vivo murine ear skin model, we demonstrated delivery of ova from LbL films into barrierdisrupted skin, uptake of the protein by skin-resident antigen-presenting cells (Langerhans cells), and transport of the antigen to the skin-draining lymph nodes. Dual incorporation of ova and CpG oligonucleotides into the nanolayers of LbL films enabled dual release of the antigen and adjuvant with distinct kinetics for each component; ova was rapidly released while CpG was released in a relatively sustained manner. Applied as skin patches, these films delivered ova and CpG to Langerhans Cells in the skin. To our knowledge, this is the first demonstration of LbL films applied for the delivery of biomolecules into skin. This approach provides a new route for storage of vaccines and other immunotherapeutics in a solid-state thin film for subsequent delivery into the immunologically-rich milieu of the skin.

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Section: Resultsmentioning

confidence: 99%

Layer-by-Layer-Assembled Multilayer Films for Transcutaneous Drug and Vaccine Delivery

Su¹,

Kim²,

Kim³

et al. 2009

ACS Nano

156

120

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“…11 Kwon et al and Murthy et al prepared a responsive polymer microsphere that degraded in the acidic lysosome to release the antigen. 12,13 Antigen cross-presentation was considered essential for the efficacy of therapeutic vaccines.…”

Section: Introductionmentioning

confidence: 99%

Improved antigen cross-presentation by polyethyleneimine-based nanoparticles

Chen¹,

Li²,

Huang³

et al. 2011

IJN

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Purpose In the development of therapeutic vaccines against cancer, it is important to design strategies for antigen cross-presentation to stimulate cell-mediated immune responses against tumor antigens. Methods We developed a polyethyleneimine (PEI)-based protein antigen delivery system to promote cross-presentation through the major histocompatibility complex (MHC) I pathway using ovalbumin (OVA) as a model antigen. PEIs formed nanoparticles with OVA by electrostatic interactions, as demonstrated by electrophoresis analysis, scanning electron microscopy, and photon correlation spectroscopy analysis. Results The nanoparticles were used to stimulate mouse bone marrow-derived dendritic cells in vitro and resulted in significantly more OVA 257–264 /MHC I complex presentation on dendritic cell surfaces. The activated dendritic cells interacted specifically with RF33.70 to stimulate interleukin-2 secretion. The cross-presentation promoting effect was more prominent in dendritic cells that had been cultured for longer periods of time (13 days). Further studies comparing the antigen presentation efficacies by other polyanionic agents, such as PLL or lysosomotropic agents, suggested that the unique “proton sponge effect” of PEI facilitated antigen escape from the endosome toward the MHC I pathway. Conclusion Such a PEI-based nanoparticle system may have the potential to be developed into an effective therapeutic vaccine delivery system.

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“…vitro. 174 By increasing the hydrophilicity of the labile crosslinker structure the loading efficiency and the resulting antigen presentation levels could be dramatically enhanced.…”

Section: Microgels Containing Hydrolytically Cleavable Crosslinkersmentioning

confidence: 99%

Light-Sensitive Polymeric Nanoparticles Based on Photo-Cleavable Chromophores

Klinger

2013

Springer Theses

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This thesis focuses on the design, synthesis and characterization of novel photo-sensitive microgels and nanoparticles as potential materials for the loading and light-triggered release/accessibility of functional compounds. In order to realize this concept, different approaches to irradiation-dependent response mechanisms have been investigated.Novel light-cleavable divinyl functionalized monomeric crosslinkers based on o-nitrobenzyl derivatives were synthesized and used for the preparation of either PMMA or hydroxyl functionalized PHEMA microgels swellable and degradable in organic solvents. By the introduction of anionic MAA moieties into the PHEMA microgels, this concept was successfully transferred to double stimuli-responsive p(HEMA-co-MAA) hydrogel nanoparticles exhibiting a pH-dependent swelling and light-induced degradation behavior in aqueous media. This sensitivity to two orthogonal triggers is proposed to combine a pHinduced post-formation loading mechanism with a pH-and light-triggered release. In addition, a new class of enzyme-and light-sensitive PAAm microgels based on (meth-)acrylate functionalized dextrans as photo-and enzymatically degradable macromolecular crosslinkers was developed by introducing a photo-labile linker between the enzymatically degradable dextran chain and the polymerizable vinyl moieties. Here, the water solubility of the crosslinkers is assumed to enable an in situ loading approach of hydrophilic compounds.A different approach to the loading of microgels is based on photo-labile covalent microgel-drug conjugates. The first step to the realization of this concept was achieved by the development of a novel functional monomer consisting of a doxorubicin molecule covalently attached to a radically polymerizable methacrylate group via a photo-cleavable linker.Moreover, in order to enable the release of hydrophobic substances in aqueous media, hydrophobic nanoparticles consisting of a photo-resist polymer were designed to be degradable upon irradiation in water. Light-triggered conversion of the initial hydrophobic polymer into hydrophilic PMAA induced in situ particle dissolution and release of nile red.Since the introduction of a stimuli-responsive shell around a microgel is assumed to prevent leakage of embedded compounds, studies on non-stimuli-responsive shell formation around preformed microgel seed particles and the formation of microgel cores in polymeric shells were conducted to investigate potential synthetic pathways to this approach.In a last attempt, the surface of PHEMA and p(HEMA-co-MAA) microgels was functionalized with cinnamoyl groups in order to trigger the (reversible) particles interaction with each other by light-induced [2+2] cycloaddition reactions.

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A macromolecular delivery vehicle for protein-based vaccines: Acid-degradable protein-loaded microgels

Cited by 389 publications

References 23 publications

Layer-by-Layer-Assembled Multilayer Films for Transcutaneous Drug and Vaccine Delivery

Layer-by-Layer-Assembled Multilayer Films for Transcutaneous Drug and Vaccine Delivery

Improved antigen cross-presentation by polyethyleneimine-based nanoparticles

Light-Sensitive Polymeric Nanoparticles Based on Photo-Cleavable Chromophores

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