The Experts below are selected from a list of 273 Experts worldwide ranked by ideXlab platform
A Kondo - One of the best experts on this subject based on the ideXlab platform.
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Affinity selection of target cells from cell surface displayed libraries: a novel procedure using Thermo-Responsive magnetic nanoparticles.
Applied microbiology and biotechnology, 2003Co-Authors: H Furukawa, R Shimojyo, N Ohnishi, H Fukuda, A KondoAbstract:Biotinylated Thermo-Responsive magnetic nanoparticles for use in affinity selection from yeast cell surface display libraries were prepared by coating magnetite nanoparticles with a Thermo-Responsive Polymer consisting of N-isopropyl acrylamide and a biotin derivative. These particles showed a reversible transition between flocculation and dispersion at around the lower critical solution temperature of 30 degrees C, above which the flocculated particles--which absorbed a large amount of avidin due to their large surface area--were quickly separable by magnet. The model library was constructed by mixing control yeast cells with target yeast cells co-displaying IgG binding protein (ZZ) and enhanced green fluorescence protein. Biotinylated IgG and avidin were subsequently added to the model library, and target cells were efficiently enriched with the biotinylated magnetic nanoparticles by avidin-biotin sandwich and ZZ-IgG interaction. The few target cells (0.001%) in the model library were enriched by up to 100% in only 5 days by an affinity selection procedure repeated four times. This novel method based on magnetic nanoparticles and a yeast cell surface display system could fulfill a wide range of applications in the analysis of protein-protein interactions and rapid isolation of novel biomolecules.
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Affinity selection of target cells from cell surface displayed libraries: a novel procedure using Thermo-Responsive magnetic nanoparticles.
Applied Microbiology and Biotechnology, 2003Co-Authors: H Furukawa, R Shimojyo, N Ohnishi, H Fukuda, A KondoAbstract:Biotinylated Thermo-Responsive magnetic nanoparticles for use in affinity selection from yeast cell surface display libraries were prepared by coating magnetite nanoparticles with a Thermo-Responsive Polymer consisting of N-isopropyl acrylamide and a biotin derivative. These particles showed a reversible transition between flocculation and dispersion at around the lower critical solution temperature of 30°C, above which the flocculated particles—which absorbed a large amount of avidin due to their large surface area—were quickly separable by magnet. The model library was constructed by mixing control yeast cells with target yeast cells co-displaying IgG binding protein (ZZ) and enhanced green fluorescence protein. Biotinylated IgG and avidin were subsequently added to the model library, and target cells were efficiently enriched with the biotinylated magnetic nanoparticles by avidin-biotin sandwich and ZZ-IgG interaction. The few target cells (0.001%) in the model library were enriched by up to 100% in only 5 days by an affinity selection procedure repeated four times. This novel method based on magnetic nanoparticles and a yeast cell surface display system could fulfill a wide range of applications in the analysis of protein-protein interactions and rapid isolation of novel biomolecules.
Xiaopeng Pei - One of the best experts on this subject based on the ideXlab platform.
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Polymer brush graft modified starch based nanoparticles as pickering emulsifiers
Langmuir, 2019Co-Authors: Xiaopeng Pei, Kankan Zhai, Chao Wang, Yukun Deng, Ying Tan, Baichao Zhang, Yungang BaiAbstract:We study biosourced core–shell particles with a starch-based core and Thermo-Responsive Polymer brush shell using surface-initiated single-electron transfer living radical Polymerization (SI-SET-LR...
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Polymer Brush Graft-Modified Starch-Based Nanoparticles as Pickering Emulsifiers
2019Co-Authors: Xiaopeng Pei, Kankan Zhai, Chao Wang, Yukun Deng, Ying Tan, Baichao Zhang, Yungang Bai, Pixin WangAbstract:We study biosourced core–shell particles with a starch-based core and Thermo-Responsive Polymer brush shell using surface-initiated single-electron transfer living radical Polymerization (SI-SET-LRP) as a Pickering stabilizer. The shell endows the Pickering stabilizer with reversible emulsification/demulsification of oil and water properties. The initiator attached to the starch-based nanosphere (Br-SNP) core particle was first fabricated using the precipitation method. Subsequently, dense poly(N-isopropylacrylamide) (PNIPAM) brush graft-modified starch-based nanoparticles (SNP-g-PNIPAM) were obtained via the SI-SET-LRP process. Interfacial properties of the resultant particles were analyzed by interfacial tensiometer measurements, as were the effects of the grafted Polymer chain length and temperature on the interfacial activity. Pickering emulsion was obtained using SNP-g-PNIPAM particles as the stabilizer. The effect of the concentration of the Pickering stabilizer on the size of emulsion droplets was analyzed. The emulsification/demulsification process of the Pickering emulsion can be reversed and easily repeated by changing the temperature
Teruo Okano - One of the best experts on this subject based on the ideXlab platform.
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Preparation of Thermo-Responsive Polymer brushes on hydrophilic Polymeric beads by surface-initiated atom transfer radical Polymerization for a highly resolutive separation of peptides
Journal of Chromatography A, 2010Co-Authors: Aya Mizutani, Yoshikatsu Akiyama, Masahiko Annaka, Jun Kobayashi, Kenichi Nagase, Akihiko Kikuchi, Hideko Kanazawa, Teruo OkanoAbstract:Abstract Poly(N-isopropylacrylamide-co-N-tert-butylacrylamide) [P(IPAAm-co-tBAAm)] brushes were prepared on poly(hydroxy methacrylate) (PHMA) [hydrolyzed poly(glycidyl methacrylate-co-ethylene glycol dimethacrylate)] beads having large pores by surface-initiated atom transfer radical Polymerization (ATRP) and applied to the stationary phases of Thermo-Responsive chromatography. Optimized amount of coPolymer brushes grafted PHMA beads were able to separate peptides and proteins with narrow peaks and a high resolution. The beads were found to have a specific surface area of 43.0 m2/g by nitrogen gas adsorption method. CoPolymer brush of P(IPAAm-co-tBAAm) grafted PHMA beads improved the stationary phase of Thermo-Responsive chromatography for the all-aqueous separation of peptides and proteins.
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thermo responsive Polymer brush grafted porous polystyrene beads for all aqueous chromatography
Journal of Chromatography A, 2010Co-Authors: Aya Mizutani, Yoshikatsu Akiyama, Masahiko Annaka, Jun Kobayashi, Kenichi Nagase, Akihiko Kikuchi, Hideko Kanazawa, Teruo OkanoAbstract:Poly(N-isopropylacrylamide) (PIPAAm) brush-grafted porous polystyrene beads with variable grafted Polymer densities were prepared using surface-initiated atom transfer radical Polymerization (ATRP) for applications in Thermo-Responsive chromatography. Utilization of these grafted beads as a stationary phase in aqueous chromatographic analysis of insulin provides a graft density-dependent analyte retention behavior. The separations calibration curve on PIPAAm-grafted polystyrene was obtained using pullulan standards and exhibited inflection points attributed to analyte diffusion into bead pores and partitioning into grafted PIPAAm brush surfaces. Presence of these inflection points supports a separation mechanism where insulin penetrates pores in polystyrene beads and hydrophobically interacts with PIPAAm brushes grafted within the pores. Control of PIPAAm brush graft density on polystyrene facilitates effective aqueous phase separation of peptides based on thermally modulated hydrophobic interactions with grafted PIPAAm within stationary phase pores. These results indicated that PIPAAm brush-grafted porous polystyrene beads prepared by surface-initiated ATRP was effective stationary phase of Thermo-Responsive chromatography for aqueous phase peptide separations.
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temperature responsive stationary phase utilizing a Polymer of proline derivative for hydrophobic interaction chromatography using an aqueous mobile phase
Journal of Chromatography A, 2006Co-Authors: Hideko Kanazawa, Akihiko Kikuchi, Eri Ayano, Chikako Sakamoto, Reiko Yoda, Teruo OkanoAbstract:A new method of chromatography is proposed, utilizing a Thermo-Responsive Polymer carrying an amino acid ester residue for the stationary phase of high-performance liquid chromatography (HPLC). We have been investigating the new concept of chromatography, a temperature-responsive chromatography, using temperature-responsive poly(N-isopropylacrylamide) (PNIPAAm)-modified surface for HPLC with a constant aqueous media as the mobile phase. In this study, we designed and synthesized Thermo-Responsive poly(acryloyl-L-proline methyl ester) and its coPolymer with N-isopropylacrylamide (NIPAAm). HomoPolymers of acryloyl-L-proline methyl ester and coPolymer were prepared by the reaction of radical telomerization. These Polymers underwent a reversible phase transition from water-soluble forms into aggregates by changing the temperature, similar to PNIPAAm. The surface properties and functions of stationary phases modified with poly(acryloyl-L-proline methyl ester) were controlled by the external temperature. In the chromatographic system, we separated steroids and amino acids with a variety of hydrophobicities using a sole aqueous mobile phase. In contrast to a PNIPAAm-modified surface, a poly(acryloyl-L-proline methyl ester)-modified surface showed a greater affinity for hydrophobic amino acids.
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thermo responsive culture dishes allow the intact harvest of multilayered keratinocyte sheets without dispase by reducing temperature
Tissue Engineering, 2001Co-Authors: Masayuki Yamato, Akihiko Kikuchi, Mika Utsumi, Ai Kushida, Chie Konno, Teruo OkanoAbstract:To develop new technology for harvesting transplantable cultured epithelium without dispase treatment, human keratinocytes were plated on culture dishes grafted with a Thermo-Responsive Polymer, poly(N-isopropylacrylamide). The grafted dish surfaces are slightly hydrophobic above 32°C, but reversibly change to hydrophilic below this temperature. According to the method of Rheinwald and Green, keratinocytes proliferated and made a multilayer on the grafted surfaces at 37°C, as on the nongrafted culture dishes. The multilayered keratinocyte sheets were detached from the grafted surfaces only by reducing temperature to 20°C without need for dispase. No cell remnants were observed on the dishes. Such cell sheet detachment was not observed on nongrafted dishes. Immunoblotting of harvested keratinocyte sheets revealed that dispase treatment disrupted E-cadherin and laminin 5, while these molecules remained intact in the keratinocyte sheets harvested by only reducing temperature from the grafted dishes. Transmis...
J Kim - One of the best experts on this subject based on the ideXlab platform.
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thermo responsive Polymer capsules in real time one step rt pcr for highly multiplex rna analysis
Advanced Healthcare Materials, 2020Co-Authors: J Kim, Seung Won Jung, Mi Yeon Kim, Bong Kyun Kim, Soon Hwan Kwon, Sang Kyung KimAbstract:Rapid and simple detection of RNA targets is in high demand due to the growing threat of pandemic viruses. One-step real-time, reverse transcription-Polymerase chain reaction (One-step RT-qPCR) using a controlled release system of Thermo-Responsive materials is developed in this paper to enable high-fidelity RNA analysis as suppressing by-products. The nanocapsules, consisting of upper critical solution temperature (UCST) material and PCR primers, carry or release the primers depending upon the temperature. The UCST nanocapsules are introduced into hydrogel microparticles incorporated with RT primers and then the target RNA is selectively amplified in the microparticle through one-step RT-qPCR. Severe side products are sharply subdued by separating the PCR primers from the RT process by means of the microparticles with nanocapsules. Because the one-step assay is now implemented in a single microparticle, multiple target RNAs can be analyzed in a simple RT-qPCR of multiple particles. Reliable 18-plex one-step RT-qPCR is successfully conducted within 30 min using single-color fluorescent optics. This work also explains the facile fabrication processes used for the Thermo-Responsive nanocapsules and hydrogel microparticles by the blending Polymerization method. Extensible multiplex analysis of influenza virus demonstrates the versatile uses of this one-step RT-qPCR platform.
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Thermo‐Responsive Polymer Capsules in Real‐Time One‐Step RT‐PCR for Highly Multiplex RNA Analysis
Advanced Healthcare Materials, 2020Co-Authors: J Kim, Seung Won Jung, Mi Yeon Kim, Bong Kyun Kim, Soon Hwan Kwon, Sang Kyung KimAbstract:Rapid and simple detection of RNA targets is in high demand due to the growing threat of pandemic viruses. One-step real-time, reverse transcription-Polymerase chain reaction (One-step RT-qPCR) using a controlled release system of Thermo-Responsive materials is developed in this paper to enable high-fidelity RNA analysis as suppressing by-products. The nanocapsules, consisting of upper critical solution temperature (UCST) material and PCR primers, carry or release the primers depending upon the temperature. The UCST nanocapsules are introduced into hydrogel microparticles incorporated with RT primers and then the target RNA is selectively amplified in the microparticle through one-step RT-qPCR. Severe side products are sharply subdued by separating the PCR primers from the RT process by means of the microparticles with nanocapsules. Because the one-step assay is now implemented in a single microparticle, multiple target RNAs can be analyzed in a simple RT-qPCR of multiple particles. Reliable 18-plex one-step RT-qPCR is successfully conducted within 30 min using single-color fluorescent optics. This work also explains the facile fabrication processes used for the Thermo-Responsive nanocapsules and hydrogel microparticles by the blending Polymerization method. Extensible multiplex analysis of influenza virus demonstrates the versatile uses of this one-step RT-qPCR platform.
Sang Kyung Kim - One of the best experts on this subject based on the ideXlab platform.
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thermo responsive Polymer capsules in real time one step rt pcr for highly multiplex rna analysis
Advanced Healthcare Materials, 2020Co-Authors: J Kim, Seung Won Jung, Mi Yeon Kim, Bong Kyun Kim, Soon Hwan Kwon, Sang Kyung KimAbstract:Rapid and simple detection of RNA targets is in high demand due to the growing threat of pandemic viruses. One-step real-time, reverse transcription-Polymerase chain reaction (One-step RT-qPCR) using a controlled release system of Thermo-Responsive materials is developed in this paper to enable high-fidelity RNA analysis as suppressing by-products. The nanocapsules, consisting of upper critical solution temperature (UCST) material and PCR primers, carry or release the primers depending upon the temperature. The UCST nanocapsules are introduced into hydrogel microparticles incorporated with RT primers and then the target RNA is selectively amplified in the microparticle through one-step RT-qPCR. Severe side products are sharply subdued by separating the PCR primers from the RT process by means of the microparticles with nanocapsules. Because the one-step assay is now implemented in a single microparticle, multiple target RNAs can be analyzed in a simple RT-qPCR of multiple particles. Reliable 18-plex one-step RT-qPCR is successfully conducted within 30 min using single-color fluorescent optics. This work also explains the facile fabrication processes used for the Thermo-Responsive nanocapsules and hydrogel microparticles by the blending Polymerization method. Extensible multiplex analysis of influenza virus demonstrates the versatile uses of this one-step RT-qPCR platform.
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Thermo‐Responsive Polymer Capsules in Real‐Time One‐Step RT‐PCR for Highly Multiplex RNA Analysis
Advanced Healthcare Materials, 2020Co-Authors: J Kim, Seung Won Jung, Mi Yeon Kim, Bong Kyun Kim, Soon Hwan Kwon, Sang Kyung KimAbstract:Rapid and simple detection of RNA targets is in high demand due to the growing threat of pandemic viruses. One-step real-time, reverse transcription-Polymerase chain reaction (One-step RT-qPCR) using a controlled release system of Thermo-Responsive materials is developed in this paper to enable high-fidelity RNA analysis as suppressing by-products. The nanocapsules, consisting of upper critical solution temperature (UCST) material and PCR primers, carry or release the primers depending upon the temperature. The UCST nanocapsules are introduced into hydrogel microparticles incorporated with RT primers and then the target RNA is selectively amplified in the microparticle through one-step RT-qPCR. Severe side products are sharply subdued by separating the PCR primers from the RT process by means of the microparticles with nanocapsules. Because the one-step assay is now implemented in a single microparticle, multiple target RNAs can be analyzed in a simple RT-qPCR of multiple particles. Reliable 18-plex one-step RT-qPCR is successfully conducted within 30 min using single-color fluorescent optics. This work also explains the facile fabrication processes used for the Thermo-Responsive nanocapsules and hydrogel microparticles by the blending Polymerization method. Extensible multiplex analysis of influenza virus demonstrates the versatile uses of this one-step RT-qPCR platform.