Silica Nanoparticles
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1 custom made Silica Nanoparticles Nano Bio Tech Nano Bio Tech
2 CUSTOM DESIGN OF INNOVATIVE NANOPARTICLE-BASED TOOLS OUR TECHNOLOGY Aczon Silica NanoParticles are made up of spherical atomic or molecular clusters, with diameter between 5 and 100 nm, and consist of two structural compartments: Core and Shell. They are made up of a Silicon-based polymer bonded to a surfactant molecule. On the shell, NPs are functionalized with the conventional functional groups (amino, carboxyl, etc.), or activated, for bioconjugations by means of a crosslinker (maleimide); the core may be left empty or it may contain fluorochromes or other molecules (linked, or otherwise, with a covalent bond) for biotechnological or other kind of applications. NanoParticles are used in the most extensive range of applications (diagnostic and clinical applications, but also in the fields of biotechnology, chemistry, pharmacy, agriculture, environment and other). HOW THEY ARE MADE ACZON CUSTOM MADE: designed for customer needs The method used to produce Aczon NanoParticles is called the Micelle - Assisted Method, which that includes the following phases (described and schematized in brief): 1. The surfactant, the silanized PEG and/or Amino silanized PEG (for functionalized NPs) are dissolved in water where they spontaneously form micelles. 2. The silica precursor is hydrolyzed and condensed extensively until the structures lock. 3. Purification is performed by means of dialysis and filtration. Fig. Micelle-assisted Method. Main Features total standard diameter of NPs: 20-25nm core diameter (including molecules): 10 nm high solubility in aqueous solutions remarkable monodispersity and high chemical stability efficient spectral performances Main Advantages variable controlled size highly reproducible and atoxic synthesis low handling of molecules linked in the Core outstanding chemical stability and atoxicity excellent monodispersity
3 CUSTOM MADE FOR IN VITRO DIAGNOSTICS Recent years have unprecedented growth of research and applications in the area of nanoscience and nanotechnology. Anticipated applications in medicine include drug delivery, both in vitro and in vivo diagnostics, nutraceuticals and production of improved biocompatible materials. Engineered nanoparticles are an important tool to realize a number of these applications (A.4). To provide a new nanomedicine platform for in vitro diagnostics, imaging, detection and treatment, as well as building blocks for multifunctional materials and different applications (A.2), it is extremely important to avail of nanoparticles with variable controlled size. Each field of application requires specific specifications; shape and dimensions are of great interest. A wide range of molecules/biomoieties can be conjugated to the nanoparticles including low molecular weight ligands (folic acid, thiamine, dimercaptosuccinic acid), peptides (RGD, LHRD, antigenic peptides, internalization peptides), proteins (BSA, transferrin, antibodies, lectins, cytokines, fibrinogen, thrombin), polysaccharides (hyaluronic acid, chitosan, dextran, oligosaccharides, heparin), polyunsaturated fatty acids (palmitic acid, phospholipids), DNA, plasmids, sirna, mirna, and so forth (A.6). The figure below shows the conjugation of nanoparticles with antibodies; it combines the properties of the nanoparticles themselves with the specific and selective recognition ability of the antibodies toward antigens. Also, the improvement in the cellular uptake as well as the major intracellular stability may be two of the major advantages of using antibody conjugated nanoparticles (A.6) What antibodies can offer NPs and what NPs can offer to Abs Antibodies Nanoparticles References A - Custom made for in vitro diagnostics: 1- Smith J. E. et al., Bioconjugated silica-coated nanoparticles for bioseparation and bioanalysis. Trends in Anal.Chem., Huo Q.S. et al., A new class of silica cross-linked micellar core-shell nanoparticles, JACS, Chan C. P. et al., New Trends in Immunoassays Adv Biochem Engin/ Biotechnol, De Jong W.H. et al., Drug delivery and nanoparticles: Applications and hazards, Int. J. of Nanomedicine, Chan C. P. et al., In vitro diagnostic prospects of nanoparticles Clin. Chim.ACTA, Arruebo M. et al., Antibody- Conjugated Nanoparticles for Biomedical Applications, J.Nanomat., Specificity Nano-size Not Specific Biological component Recruits components of the immune system Can find specific target Specific Carriers Can activate the immune system Several proprieties Thrmal, Magnetics, Imaging Drug delivery-controlled Many properties Versatility Fig. Advantages of conjugating nanoparticles with antibodies (Arruebo M. et al., Antibody- Conjugated Nanoparticles for Biomedical Applications, J.Nanomat., 2009). Why Aczon NanoParticles for in vitro diagnostics? Aczon NanoParticles produced according to customer specifications are created by including fluorescent and non-fluorescent molecules, functionalized with different chemical groups and/or conjugated with proteins (such as antibodies, or enzymes), peptides, nucleotide sequences and much more. We can synthesize NPs incorporating special molecules of the customer and conjugate or functionalize according to specific requests.
4 CUSTOM MADE FOR IN VIVO DIAGNOSIS: NanoTrack TM The development of biomedical imaging techniques, such as computed X-ray tomography (CT), optical imaging, and magnetic resonance imaging (MRI), has brought significant advances for diagnosis and therapy. Inorganic nanoparticles are emerging as promising probes to shed light on biological processes and diseases occurring at molecular and cellular levels. References B - Custom made for in vivo diagnosis: 1- Young-wook J. et al., Chemical Design of Nanoparticle Probes for High-Performance Magnetic Resonance Imaging, Angew. Chem. Int., Gagnon M.K.J. et al. Highthroughput in vivo screening of targeted molecular imaging agents PNAS., Perrault S.D. et al. In vivo assembly of nanoparticles components to improve targeted cancer imaging PNAS., Pantazis P. Second Harmonic generating (SHG) nanoprobes for in vivo imaging PNAS., Hwang D. W. et al., Molecular Imaging Using PET/MRI Particle, The Open Nuclear Med. J., They have the potential to advance imaging from its current anatomy-based level to the molecular level, so-called molecular imaging. Upon conjugation with target-specific biomolecules, these tiny probes (1 100nm) can travel through the human body in the blood and lymphatic vessels and they can identify the desired target through specific biological interactions, such as antibody antigen, nucleic acid hybridization, and gene expression (B.1). The development of efficient imaging probes has accelerated the possibility to detect the grafted cell migration in vivo and evaluate drug response in the molecular imaging field, offering repetitive imaging detection, signal quantification and tomographic capabilities. Furthermore, the development of multimodal imaging nanoprobes can present a multidisciplinary approach for developing merged PET-MRI imaging devices, drug delivery and cancer tracking (B.5). The figure below shows a schematic nanoparticles assembly with contrast agent in vivo. Gold nanoparticles stabilized with biotinylated PEG (denoted as biotin-peg anchor) are injected as a first step. These enter tumors through leaky vasculature and passively accumulate in the extracellular matrix over 24 h. Fluorescently labelled streptavidin is injected, which leaks into tumors and interacts with biotin on the gold nanoparticles in the interstitium. This favourably alters the contrast agent s tumor accumulation kinetics (B.3). Fig. Schematic nanoparticle assembling with contrast agent in vivo (Perrault S.D. et al. In vivo assembly of nanoparticles components to improve targeted cancer imaging PNAS., 2010). Why Aczon NanoParticles for in vivo diagnosis? Aczon NanoParticles fit into this context and are designed for diagnostic applications, instead of and in support of the conventional diagnostic imaging technologies. NanoParticles are dull and consequently visible to conventional In Vivo Diagnostic technologies (NMR, PET, ). Moreover, thanks to their interesting features, increased image resolution, absence of cytotoxicity and, in particular, their high diffusion capacity and their variable controlled size, they can enhance permeability and retention at cancer cells level. According to specific customer needs, we can design and produce fluorescent or magnetic NanoParticles to be used as contrast agents and/or tracers.
5 CUSTOM MADE FOR PHARMA APPLICATIONS: NanoCarriers TM The use of nanotechnology in medicine and, more specifically drug delivery, is set to spread rapidly. Currently many substances are being investigated for drug delivery and more specifically for cancer therapy. Interestingly, pharmaceutical sciences are using nanoparticles to reduce toxicity and side effects of drugs. The reason why these nanoparticles are attractive for medical purposes is due to their important and unique features, such as their surface to mass ratio, which is much larger than that of other particles, their quantum properties and their ability to adsorb and carry other compounds. NanoParticles consist of at least two components, one of which is a pharmaceutically active ingredient. The primary goals for research of nanobio-technologies in drug delivery include: more specific drug targeting and delivery, reduction in toxicity while maintaining therapeutic effects, greater safety and biocompatibility, faster development of new safe medicine (C.1), and extended circulation half-life and improved therapeutic index (C.2). The figure below highlights the various surface modifications that are commonly pre-engineered, such as cellular targeting, particle stealthing and organelle targeting. Ligands to extend circulation half-life and to reduce immunogenicity (usually polyethylene glycol (PEG) chains) are linked to the surface of the nanoparticle together with ligands to promote targeting. These ligands can be antibodies, aptamers or small molecules known to bind to surface proteins expressed on target cells or that are capable of guiding particle localization once inside the cell. Chemotherapeutics or other biologically relevant cargo are encapsulated inside the nanoparticle. Release of the cargo at the intended site of action is typically achieved through the incorporation of a stimuli-responsive material that changes state upon exposure to the targeted environment (C.2). References C - Custom made for pharma applications: 1- De Jong W.H. et al., Drug delivery and nanoparticles: Applications and hazards, Int. J. of Nanomedicine, Petros R.A. et al., Strategies in the design of nanoparticles for therapeutic applications, Nature Reviews Drug Discovery, Luo J. et al, Size-Tunable, Multifunctional Micelles for Efficient Paclitaxel Delivery for Cancer Treatment, Bioconugate. Chem., Abhimanyu S. et al., Harnessing structure-activity relationship to engineer a cisplatin nanoparticle for enhanced antitumor efficacy, PNAS, Tennant D.A. et al., Targeting metabolic transformation for cancer therapy, Nature Reviews, Fig. Schematic representation of an engineered nanoparticle (Petros R.A. et al., Strategies in the design of nanoparticles for therapeutic applications, Nature Reviews Drug Discovery, 2010). Why Aczon NanoParticles for Pharma applications? Aczon NanoParticles, synthesized with specific biomolecules, are very attractive for drug delivery applications, because they are innovative tools that offer distinctive features and advantages (high diffusion capacity, easier to reach the target cells, considerable concentration of the active principle, effective concentration in vivo and increase local drug concentration). We can incorporate drugs or other cytotoxic molecules and/or synthesize NPs with specific bio-molecules that are of great interest to researchers and manufacturers.
6 CUSTOM MADE FOR ENVIRONMENTAL APPLICATIONS Sensing of biological agents, diseases, and toxic materials is an important goal for biomedical diagnosis, forensic analysis, and environmental monitoring. The unique physicochemical properties of NPs coupled with the inherent increase in signal-to-noise ratio provided by miniaturization makes these systems promising candidates for sensing applications. The integration of nanoparticles with biomolecules in the area of biosensing has implemented different subset of environmental applications: colorimetric sensing, fluorescence sensing, chemical and electrochemical sensing. References D - Custom made for environmental applications: 1- Zhao, X. J. et al., A rapid bioassay for single bacterial cell quantitation using bioconjugated nanoparticles. Proc. Nat. Acad. Sci., Tan W. et al, Bionanotechnology Based on SilicaNanoparticles, Medicinal Research Reviews, Zhang, Q. et al, Microbial detection in microfluidic devices through dual staining of quantum dots-labeled immunoassay and RNA hybridization, Anal. Chim. Acta, Liu W., Nanoparticles and their biological and environment applications, J. of Biosci. Bioeng., De M. et al., Applications of Nanoparticles in Biology, Adv. Mater., Fig. Images of bacterial cells. (A) SEM image of E. coli O157:H7 cell incubated with Ab-NPs. (B) SEM image of E. coli DH5α cell (negative control) incubated with Ab-NPs for E. coli O157:H7. (C) Fluorescence image of E. coli O157:H7 after incubation with Ab-NPs. The fluorescence intensity is strong, enabling single-bacterium cell identification in aqueous solution (Zhao X. J. et al., PNAS, 2004). As example of colorimetric sensing, gold nanoparticles exhibit unique optical and electronic properties based on size and shape. They show an intense absorption peak from 500 to 550 nm arising from surface plasmon resonance (SPR). The SPR band is sensitive to the surrounding environment, signaling changes in solvent and binding: a particularly useful output is the red-shift (to ca. 650 nm). This phenomenon leads to the popular and widely applicable colorimetric sensing (D.5). The detection and monitoring of microoganisms can be further accelerated using nanoparticles in a fluorescence labeling system, such as microfluidic devices, nanoparticle-based fluorescence reporting systems and others (D.3). Antibodies against antigens, specific for E. coli strain O157, were conjugated with fluorescent silica nanoparticles and used in immunological assay to achieve rapid bacterial detection at the single-cell level (as shown in the figure below). With the improvement in the fluorescence reporting system, the fluorescence intensity emitted by one E. coli O157 cell was sufficient to be detected using a normal spectrofluorometer or to be accurately enumerated using a flow cytometer (D.2, D.4). Several studies of chemical sensing have utilized bimetallic nanoparticles as an effective oxidant in the cleanup of environmental contaminants, mainly because nanoparticles can diffuse or penetrate into a contamination zone that microparticles are unable to reach (D.4). Why Aczon NanoParticles for Environmental applications? Aczon team has outstanding expertise in organic and inorganic synthesis of molecules and NanoParticles doped and/or functionalized with different chemical groups and crosslinkers or conjugated with bio-molecules, so we can synthesize different kinds of NPs: especially Silica NanoParticles (proprietary technology); alternatively, to better meet the needs of customers, we can offer and suggest other ones (such as gold and many other). We can incorporate special molecules of the customer and conjugate or functionalize NPs according to specific requests.
7 CUSTOM MADE GUIDELINES 1- Type of NanoParticles (Silica, Gold, other). 2- NanoParticles size: Silica (range 5-100nm), Gold (range nm), other (if possible). 3- Core: fluorescent molecules (different fluorophores with specific absorbance and emission), small drugs (Doxorubicin, Cisplatin, other drug molecules or a mix of them) and other chemical compounds. 4- Shell: functionalization with different chemical groups (such as -NH2, COOH, maleimide, -N3, -SH and others if possible). 5- Bio-conjugation: conjugation with protein (such as antibodies), peptides, nucleic acids and other bio or chemical molecules. 6- Special requirements: buffer, concentration, solubility and others. 7- Testing requirements: standard test, endotoxin test, other test required if possible. NanoParticles size Testing requirements Type of NanoParticles Shell Core Core Bio-conjugation Special requirements ACZON CUSTOM MADE: designed for customer needs The guidelines in tangram system : a great project for ACZON Custom Made.
8 Plan Feedback Develop Deliver Act ACZON CUSTOM MADE: designed for customer needs Fig. Schematic representation of synthesized NPs incorporating the customer s molecules, and conjugated or functionalized according to specific requests. PLAN DEVELOP ACT - Establish customer needs (applications/ conventional technology problems ) - Prepare a preliminary definition of the product s specifications and performances in collaboration with your scientific consultant - Prepare a feasibility study and design the project - Project approval - Produce the prototype - Validate the master plan - Assess the reagent s reproducibility, specificity, stability and ruggedness - Present the results - Customer approval/modifications - Production scale-up DELIVER - Delivery to the customer FEEDBACK - Customer s assessment of the product - Technical support from Aczon s Custom Made Team Call us or check out our website for more technical information! CAT REF NR BRCMD--0812W ACZON S.r.l. Via Lavino, 265-D Monte San Pietro - Bologna - ITALY Tel.: Fax:
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