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In JoVE (3)
- Combinatorial Synthesis of and High-throughput Protein Release from Polymer Film and Nanoparticle Libraries
- Harvesting Murine Alveolar Macrophages and Evaluating Cellular Activation Induced by Polyanhydride Nanoparticles
- High-throughput Synthesis of Carbohydrates and Functionalization of Polyanhydride Nanoparticles
Other Publications (3)
Articles by Ana V. Chavez-Santoscoy in JoVE
JoVE Bioengineering
Combinatorial Synthesis of and High-throughput Protein Release from Polymer Film and Nanoparticle Libraries
Department of Chemical and Biological Engineering, Iowa State University
This method describes the combinatorial synthesis of biodegradable polyanhydride film and nanoparticle libraries and the high-throughput detection of protein release from these libraries.
JoVE Bioengineering
Harvesting Murine Alveolar Macrophages and Evaluating Cellular Activation Induced by Polyanhydride Nanoparticles
1Department of Chemical and Biological Engineering, Iowa State University, 2Department of Veterinary Microbiology and Preventive Medicine, Iowa State University
Herein, we describe protocols for harvesting murine alveolar macrophages, which are resident innate immune cells in the lung, and examining their activation in response to co-culture with polyanhydride nanoparticles.
JoVE Bioengineering
High-throughput Synthesis of Carbohydrates and Functionalization of Polyanhydride Nanoparticles
1Department of Chemical and Biological Engineering, Iowa State University, 2Department of Chemistry, Iowa State University
In this article, a high throughput method is presented for the synthesis of oligosaccharides and their attachment to the surface of polyanhydride nanoparticles for further use in targeting specific receptors on antigen presenting cells.
Other articles by Ana V. Chavez-Santoscoy on PubMed
Prediction of Trapping Zones in an Insulator-based Dielectrophoretic Device
A mathematical model is implemented to study the performance of an insulator-based dielectrophoretic device. The geometry of the device was captured in a computational model that solves Laplace equation within an array of cylindrical insulating structures. From the mathematical model it was possible to predict the location and magnitude of the zones of dielectrophoretic trapping of microparticles. Simulation and experimental results of trapping zones are compared for different operating conditions.
Controlled Microparticle Manipulation Employing Low Frequency Alternating Electric Fields in an Array of Insulators
Low frequency alternating current insulator-based dielectrophoresis is a novel technique that allows for highly controlled manipulation of particles. By varying the shape of an AC voltage applied across a microchannel containing an array of insulating cylindrical structures it was possible to concentrate and immobilize microparticles in bands; and then, move the bands of particles to a different location. Mathematical modeling was performed to analyze the distribution of the electric field and electric field gradient as function of the shape of the AC applied potential, employing frequencies in the 0.2-1.25 Hz range. Three different signals were tested: sinusoidal, half sinusoidal and sawtooth. Experimental results demonstrated that this novel dielectrophoretic mode allows highly controlled particle manipulation.
Tailoring the Immune Response by Targeting C-type Lectin Receptors on Alveolar Macrophages Using "pathogen-like" Amphiphilic Polyanhydride Nanoparticles
C-type lectin receptors (CLRs) offer unique advantages for tailoring immune responses. Engagement of CLRs regulates antigen presenting cell (APC) activation and promotes delivery of antigens to specific intracellular compartments inside APCs for efficient processing and presentation. In these studies, we have designed an approach for targeted antigen delivery by decorating the surface of polyanhydride nanoparticles with specific carbohydrates to provide pathogen-like properties. Two conserved carbohydrate structures often found on the surface of respiratory pathogens, galactose and di-mannose, were used to functionalize the surface of polyanhydride nanoparticles and target CLRs on alveolar macrophages (AMϕ), a principle respiratory tract APC. Co-culture of functionalized nanoparticles with AMϕ significantly increased cell surface expression of MHC I and II, CD86, CD40 and the CLR CIRE over non-functionalized nanoparticles. Di-mannose and galactose functionalization also enhanced the expression of the macrophage mannose receptor (MMR) and the macrophage galactose lectin, respectively. This enhanced AMϕ activation phenotype was found to be dependent upon nanoparticle internalization. Functionalization also promoted increased AMϕ production of the pro-inflammatory cytokines IL-1β, IL-6 and TNF-α. Additional studies demonstrated the requirement of the MMR for the enhanced cellular uptake and activation provided by the di-mannose functionalized nanoparticles. Together, these data indicate that targeted engagement of MMR and other CLRs is a viable strategy for enhancing the intrinsic adjuvant properties of nanovaccine adjuvants and promoting robust pulmonary immunity.
