Biofunctionalization of upconversion nanoparticles for intracellular labeling and imaging (original) (raw)
Related papers
Surface Assembly and Intracellular Delivery of Biomolecule Conjugated Nanomaterials
2008
2.1 Introduction………………………………………………………………………….8 2.2 Methods and Results………………………………………………………….……9 2.3 Discussion and Model….…….……………….………………………………...20 3. MAINTAINING STRUCTURE AND FUNCTION OF A HISTIDINE-TAGGED PROTEIN ATTACHED TO A 1.5 nm Au NANOPARTICLE…..……………………….23 3.1 Introduction………………………….……………………………………………..23 3.2 Materials and Methods……………………………………………………………25 3.3 Results…………………………………………………………………………..….28 3.4 Conclusions…………………………………………………………..……………38 4. DEVELOPMENT OF A BIMODAL MRI CONTRAST AGENT………………………...41 4.1 Introduction…………………………………..…………………………………….41 4.2 Materials and Methods….…………...………………………..……………….….46 4.3 Results……….………..…………………………………………………………....50 4.4 Conclusions……………………….…………………………………………..…...55 4.5 Future Work……………………………………………………………….……….56 5. NANOMEDIATED DELIVERY OF DUAL GENE REGULATORS…..………………..57 5.1 Introduction………………………………………………………………………...57 5.2 Materials and Methods……………………………………………………………59 v 5.3 Results………………………………………………………………………………61 5.4 Discussion………………………………………………………………………….71 REFERENCES…………………………………………………………………………………72 BIOGRAPHICAL SKETCH……………………………………………………….…..………87 vi LIST OF TABLES 2.1 FT-IR peak assignments of His 6-peptide and His 6 peptide with AuNP………….14 2.2 FT-IR peak assignments of Cys-peptide and Cys-peptide with AuNP………..…16 2.3 FT-IR peak assignments of Ser-peptide and Ser-peptide with AuNP……………17 3.1 FITC fluorescence recovery rates with FGF1 and FGF1-heparin………………..29 4.1 T1 and T2 relaxation rates for InP/ZnS-CAAKA-DOTA-Dy(III) cell layer experiments in agarose tissue mimic……………………………………………..…52 4.2 T1, T2, T2 linked, and T2 star results for InP/ZnS-CAAKA-DOTA-Dy(III) cell dosing experiments …………………………………………………………………..53 4.3 T1 and T2 relaxation rates for InP/ZnS-CAAKATat-DOTA-Dy(III) with and without cationic liposome transfecting agent………………………………………………..55 vii LIST OF FIGURES 1.1 Crystallographic model of 12 base pair duplex DNA and DsRed fluorescent protein…………………………………………………………………………………….3 1.2 Microscope image of CHO cells and TEM of 1.5 nm AuNP……….
2018
Lanthanide ion doped upconversion nanoparticles (UCNPs) that can convert low-energy infrared photons into high-energy visible and ultraviolet photons, are becoming highly sought-after for advanced biomedical and biophotonics applications. Their unique luminescent properties enable UCNPs to be applied for diagnosis, including biolabeling, biosensing, bioimaging and multiple imaging modality, as well as therapeutic treatments including photothermal and photodynamic therapy, bio-reductive chemotherapy and drug delivery. For the employment of the inorganic nanomaterials into biological environment, it is critical to bridge the gap in between nanoparticles and biomolecules via surface modifications and subsequent functionalisation. This work reviews the various ways to surface modify and functionalise UCNPs so as to impart different functional molecular groups to the UCNPs surfaces for a board range of applications in biomedical areas. We discussed commonly used base functionalities, inc...
2011
We report a simple and rapid method to prepare extremely bright, functionalized, stable, and biocompatible conjugated polymer nanoparticles incorporating functionalized polyethylene glycol (PEG) lipids by reprecipitation. These nanoparticles retain the fundamental spectroscopic properties of conjugated polymer nanoparticles prepared without PEG lipid, but demonstrate greater hydrophilicity and quantum yield compared to unmodified conjugated polymer nanoparticles. The sizes of these nanoparticles, as determined by TEM, were 21-26 nm. Notably, these nanoparticles were prepared with several PEG lipid functional end groups, including biotin and carboxy moieties that can be easily conjugated to biomolecules. We have demonstrated the availability of these end groups for functionalization using the interaction of biotin PEG lipid conjugated polymer nanoparticles with streptavidin. Biotinylated PEG lipid conjugated polymer nanoparticles bound streptavidin-linked magnetic beads, while carboxy and methoxy PEG lipid modified nanoparticles did not. Similarly, biotinylated PEG lipid conjugated polymer nanoparticles bound streptavidin-coated glass slides and could be visualized as diffraction-limited spots, while nanoparticles without PEG lipid or with non-biotin PEG lipid end groups were not bound. To demonstrate that nanoparticle functionalization could be used for targeted labelling of specific cellular proteins, biotinylated PEG lipid conjugated polymer nanoparticles were bound to biotinylated anti-CD16/32 antibodies on J774A.1 cell surface receptors, using streptavidin as a linker. This work represents the first demonstration of targeted delivery of conjugated polymer nanoparticles and demonstrates the utility of these new nanoparticles for fluorescence based imaging and sensing.We report a simple and rapid method to prepare extremely bright, functionalized, stable, and biocompatible conjugated polymer nanoparticles incorporating functionalized polyethylene glycol (PEG) lipids by reprecipitation. These nanoparticles retain the fundamental spectroscopic properties of conjugated polymer nanoparticles prepared without PEG lipid, but demonstrate greater hydrophilicity and quantum yield compared to unmodified conjugated polymer nanoparticles. The sizes of these nanoparticles, as determined by TEM, were 21-26 nm. Notably, these nanoparticles were prepared with several PEG lipid functional end groups, including biotin and carboxy moieties that can be easily conjugated to biomolecules. We have demonstrated the availability of these end groups for functionalization using the interaction of biotin PEG lipid conjugated polymer nanoparticles with streptavidin. Biotinylated PEG lipid conjugated polymer nanoparticles bound streptavidin-linked magnetic beads, while carboxy and methoxy PEG lipid modified nanoparticles did not. Similarly, biotinylated PEG lipid conjugated polymer nanoparticles bound streptavidin-coated glass slides and could be visualized as diffraction-limited spots, while nanoparticles without PEG lipid or with non-biotin PEG lipid end groups were not bound. To demonstrate that nanoparticle functionalization could be used for targeted labelling of specific cellular proteins, biotinylated PEG lipid conjugated polymer nanoparticles were bound to biotinylated anti-CD16/32 antibodies on J774A.1 cell surface receptors, using streptavidin as a linker. This work represents the first demonstration of targeted delivery of conjugated polymer nanoparticles and demonstrates the utility of these new nanoparticles for fluorescence based imaging and sensing. Electronic supplementary information (ESI) available: Additional TEM data, supplemental light scattering measurements, absorbance and fluorescence emission spectra, and photostability measurements. See DOI: 10.1039/c0nr00746c
Nanoscale, 2012
All chemicals used in the synthesis of the nanoparticles were purchased from Sigma-Aldrich and used as received. Millipore water was used in the preparation of all aqueous solutions used in the characterization of the samples. Synthesis of Oleate-Capped-Ln 3+-UCNPs. Oleate-capped NaGdF 4 :Er 3+ 2%, Yb 3+ 20% nanoparticles (oleate-capped-Ln 3+-UCNPs) were synthesized via the thermal decomposition procedure 1, 2. In the first step, the precursors (Solution A) were prepared by mixing 0.975 mmol Gd 2 O 3 (99.99 %), 0.25 mmol Yb 2 O 3 (99.99 %), and 0.025 mmol Er 2 O 3 (99.99 %) with 5 mL trifluoroacetic acid (99 %) and 5 mL of distilled water in a 100 mL three-neck round-bottom flask. The solution was stirred and refluxed at 80 °C for 12 h or until a clear solution was obtained and the temperature was lowered to 60 °C to slowly evaporate excess trifluoroacetic acid and water. In the second step, 2.5 mmol sodium trifluoroacetic acid CF 3 COONa (98 %) was added to the dried lanthanide trifluoroacetate precursors and mixed with 7.5 mL each of oleic acid and 1-octadecene (Solution A). In a separate three-neck round bottom flask 12.5 mL each of the coordinating ligand oleic acid (90 %) and the non-coordinating solvent 1-octadecene (90 %) were added (Solution B). Both solutions were placed under vaccum at a temperature of 150 °C, degassed to remove residual water and oxygen with stirring for 30 minutes. Solution B was heated under argon flow at a rate of approximately 8 °C/min, to 310 °C. Solution A was added to Solution B using a mechanical pump system at a rate of 1.5 mL/min (Harvard Apparatus Econoflow). The solution was maintained at 310 °C and stirred vigorously for 2 h to form the oleate-capped NaGdF 4 :Er 3+ 2%, Yb 3+ 20% nanoparticles. After 2 h, the mixture was allowed to cool to room temperature, and the oleate-capped-Ln 3+-UCNPs were precipitated by the addition of hexane/ethanol (1:4 v/v) and isolated via centrifugation at 3000 rpm for 15 minutes. The resulting pellet was then washed once with ethanol and further purified by dispersing in a minimum amount of hexane and precipitated with excess ethanol. The resulting pellet was subsequently washed with acetone and isolated via centrifugation. The resulting oleate-capped-Ln 3+-UCNPs were dried.
Single-step surface functionalization of polymeric nanoparticles for targeted drug delivery
Biomaterials, 2009
Targeted drug delivery using nanocarriers is achieved by functionalizing the carrier surface with a tissue-recognition ligand. Current surface modification methods require tedious and inefficient synthesis and purification steps, and are not easily amenable to incorporating multiple functionalities on a single surface. In this report, we describe a versatile, single-step surface functionalizing technique for polymeric nanoparticles. The technique utilizes the fact that when a diblock copolymer like polylactide-polyethylene glycol (PLA-PEG) is introduced in the oil/water emulsion used in polymeric nanoparticle formulation, the PLA block partitions into the polymer containing organic phase and PEG block partitions into the aqueous phase. Removal of the organic solvent results in the formation of nanoparticles with PEG on the surface. When a PLA-PEG-ligand conjugate is used instead of PLA-PEG copolymer, this technique permits a 'one-pot' fabrication of ligandfunctionalized nanoparticles. In the current study, the IAASF approach facilitated the simultaneous incorporation of biotin and folic acid, known tumor-targeting ligands, on drug-loaded nanoparticles in a single step. Incorporation of the ligands on nanoparticles was confirmed by using NMR, surface plasmon resonance, transmission electron microscopy and tumor cell uptake studies. Simultaneous functionalization with both ligands significantly enhanced nanoparticle accumulation in tumors in vivo, and resulted in greatly improved efficacy of paclitaxel-loaded nanoparticles in a mouse xenograft tumor model. This new surface functionalization approach will enable the development of targeting strategies based on the use of multiple ligands on a single surface to target a tissue of interest.
Biomimetic Surface Engineering of Lanthanide-Doped Upconversion Nanoparticles as Versatile Bioprobes
Angewandte Chemie International Edition, 2012
A general and versatile biomimetic approach to synthesize water dispersible and functionalizable upconverting nanoparticles (UCNPs) for selective imaging of live cancer cells is reported. The approach involves coating the surface of UCNPs with a monolayer of phospholipids containing different functional groups, allowing for conjugation of many molecules for a wide range of applications in fields such as bioinspired nanoassembly, biosensing, and bio-medicine.
ACS Applied Bio Materials
Engineered polymeric nanoparticles (NPs) have been comprehensively explored as potential platforms for diagnosis and targeted therapy for several diseases including cancer. Herein, we designed functional poly(acrylic acid)-b-poly(butyl acrylate) (PAA-b-PBA) NPs using reversible addition-fragmentation chaintransfer (RAFT)-mediated emulsion polymerization via polymerization-induced self-assembly (PISA). The hydrophilic PAA-macroRAFT, forming a stabilizing shell (i.e., corona), was chainextended using the hydrophobic monomer n-butyl acrylate (n-BA), resulting in stable, monodisperse, and reproducible PAA-b-PBA NPs, typically having a diameter of 130 nm. The surface engineering of the PAA-b-PBA NP post-PISA were explored using a two-step approach. The hydrophilic NP-shell corona was modified with allyl groups under mild conditions, using allylamine in water, which resulted in stable allyl-functional NPs (allyl-NPs) suitable for further bioconjugation. The allyl-NPs were subsequently conjugated with a thiol-functional fluorescent dye (BODIPY-SH) to the allyl groups using "thiol-ene"-click chemistry, to mimic the attachment of a thiol-functional target ligand. The successful attachment of BODIPY-SH to the allyl-NPs was corroborated by UV−vis spectroscopy, showing the characteristic absorbance of the BODIPY-fluorophore at 500 nm. Despite modification of NPs with allyl groups and attachment of BODIPY-SH, the NPs retained their colloidal stability and monodispersity as indicated by DLS. This demonstrates that post-PISA functionalization is a robust method for synthesizing functional NPs. Neither the NPs nor allyl-NPs showed significant cytotoxicity toward RAW264.7 or MCF-7 cell lines, which indicates their desirable safety profile. The cellular uptake of the NPs using J774A cells in vitro was found to be time and concentration dependent. The anti-cancer drug doxorubicin was efficiently (90%) encapsulated into the PAA-b-PBA NPs during NP formation. After a small initial burst release during the first 2 h, a controlled release pattern over 7 days was observed. The present investigation demonstrates a potential method for functionalizing polymeric NP post-PISA to produce carriers designed for targeted drug delivery.
International Journal of Molecular Sciences, 2012
In vivo optical Imaging is an inexpensive and highly sensitive modality to investigate and follow up diseases like breast cancer. However, fluorescence labels and specific tracers are still works in progress to bring this promising modality into the clinical day-today use. In this study an anti-MUC-1 binding single-chain antibody fragment was screened, produced and afterwards labeled with newly designed and surface modified NaYF 4 :Yb,Er upconversion nanoparticles as fluorescence reporter constructs. The MUC-1 binding of the conjugate was examined in vitro and in vivo using modified state-of-the-art small animal Imaging equipment. Binding of the newly generated upconversion nanoparticle
Functionalized Biocompatible Nanoparticles for Site-Specific Imaging and Therapeutics
Advances in Polymer Science, 2011
The applicability of nanoparticles is determined by their unique sizedependent properties, such as their optical and magnetic properties, which make them very attractive candidates for numerous biomedical applications such as drug delivery nanosystems, diagnostic biosensors, and imaging nanoprobes for magnetic resonance imaging contrast agents. Surface chemistry defines the functional properties and biological reactivity of these nanocrystals. Targeted delivery of therapeutics has the potential to localize therapeutic agents to a specific tissue as a mechanism to enhance treatment efficacy and mitigate side effects. Moieties that combine imaging and therapeutic modalities in a single macromolecular construct may confer advantages in the development and applications of nanomedicine. Here, an insight into the development of various kinds of functionalized biocompatible nanoparticles for site-specific imaging and therapeutics is discussed in detail.
Macromolecular Rapid Communications, 2015
Here, the synthesis and the characterization of novel amphiphilic graft copolymers with tunable properties, useful in obtaining polymeric fl uorescent nanoparticles for application in imaging, are described. These copolymers are obtained by chemical conjugation of rhodamine B (RhB) moieties, polylactic acid (PLA), and O-(2-aminoethyl)-O′-methyl poly(ethylene glycol) (PEG) on α,β-poly(N-2-hydroxyethyl)-D,L-aspartamide (PHEA). In particular, PHEA is fi rst functionalized with RhB to obtain PHEA-RhB with a derivatization degree in RhB (DD RhB) equal to 0.55 mol%. By varying the reaction conditions, different amounts of PLA are grafted on PHEA-RhB to obtain PHEA-RhB-PLA with DD PLA equal to 1.9, 4.0, and 6.2 mol%. Then, PEG chains are grafted on PHEA-RhB-PLA derivatives to obtain PHEA-RhB-PLA-PEG graft copolymers. The preparation of polymeric fl uorescent nanoparticles with tunable properties and spherical shape is described by using PHEA-RhB-PLA-PEG with DD in PLA and PEG equal to 4.0 and 4.9 mol%, by following easily scaling up processes, such as emulsion-solvent evaporation and high pressure homogenization (HPH)-solvent evaporation techniques.