• Title/Summary/Keyword: Protein nanoparticle

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Protein Delivery System based on Various Polysaccharides

  • Jung, Young-Seok;Na, Kun
    • Journal of Pharmaceutical Investigation
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    • v.41 no.4
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    • pp.197-204
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    • 2011
  • Delivery of therapeutic protein drugs is a hot issue in the clinical application, because protein drugs have low side effects and highly therapeutic effects compared with chemical drugs. Despite their prominent advantages, protein drugs have high risk for human therapy such as their easy degradation by proteolytic enzymes, renal filtration and immune response. Over the past few decades, a large number of polysaccharides as vehicles for the protein delivery system have been developed to overcome the problems. This review presents the studies on protein delivery based on polysaccharides used as stabilizer and vehicles comprising nano- or microspheres to overcome inherent limitations of therapeutic proteins.

Positively Charged Silver Nanoparticles Threaded on Carbon Nanotube for the Efficient Delivery of Negatively Charged Biomolecules

  • Park, Hyung-Seok;Hwang, Ji-Young;Shin, Ueon-Sang;Kim, Hae-Won;Gong, Myoung-Seon
    • Bulletin of the Korean Chemical Society
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    • v.32 no.10
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    • pp.3581-3586
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    • 2011
  • Silver nanoparticle (Ag-NPs)-immobilized and amine-functionalized carbon nanotubes (MWCNTs), MWCNT-Ag-$NH_2$, were easily prepared in order to develop an efficient delivery system of biomolecules without complicated processes of manufacture. For this, Ag-NPs-immobilized MWCNTs, MWCNT-Ag, were initially prepared in order to create large surface area to enable more efficient linkage with guest-molecules using pristine MWCNTs. The Ag-NPs on MWCNTs were further positively functionalized with 2-aminoethanthiol to allow ionic linkage with biomolecules. Ultimately, the positively charged delivery system proved to be highly effective for the binding capacity of bovine serum albumin (BSA) as a negatively charged model protein, when compared to that of lysozyme used as a positively charged model protein. The releasing profile of BSA was observed in almost linear pattern for about two weeks in a saline solution. This study demonstrated the potential usefulness of the pristine MWCNTs in conjunction with Ag-NPs for the selective delivery of many (negatively or positively) charged biomolecules including proteins and genes.

Gel Electrophoresis Analysis of the Hard Coronas of Human Serum Albumin on Silica Nanoparticles: Size Dependence of Corona Formation

  • Kim, Sung-Jong;Han, Sang Yun
    • Bulletin of the Korean Chemical Society
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    • v.35 no.9
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    • pp.2621-2624
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    • 2014
  • The rapid and spontaneous adsorption of proteins on nanoparticle (NP) surfaces in biological fluids such as blood is an important phenomenon as it possibly determines "what the cells see" and, thus, the fates of NPs in living organisms. In order to quantitatively understand protein coronas at the molecular level, we investigated human serum albumin (HSA) coronas that were produced on silica NPs of 20 nm and 50 nm diameters using conventional gel electrophoresis. Analysis of the concentration dependence of protein adsorption showed that HSA coronas preferentially formed a monolayer on silica NPs and revealed the presence of hard protein coronas. HSA adsorption was clearly dependent on NP size, and this might be due to the different surface curvatures of NPs of different sizes.

Recent advances in utilization of photochemical internalization (PCI) for efficient nano carrier mediated drug delivery

  • Park, Wooram;Park, Sin-Jung;Lee, Jun;Na, Kun
    • Biomaterials and Biomechanics in Bioengineering
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    • v.2 no.1
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    • pp.1-13
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    • 2015
  • Despite recent progresses in nanoparticle-based drug delivery systems, there are still many unsolved limitations. Most of all, a major obstacle in current nanoparticle-based drug carrier is the lack of sufficient drug delivery into target cells due to various biological barriers, such as: extracellular matrix, endolysosomal barrier, and drug-resistance associated proteins. To circumvent these limitations, several research groups have utilized photochemical internalization (PCI), an extension of photodynamic therapy (PDT), in design of innovative and efficient nano-carriers drug delivery. This review presents an overview of a recent research on utilization of PCI in various fields including: anti-cancer therapy, protein delivery, and tissue engineering.

Optimization of Protoplast Isolation and Ribonucleoprotein/Nanoparticle Complex Formation in Lentinula edodes (표고버섯의 원형질체 분리 최적화와 RNPs/나노파티클 복합체 형성)

  • Kim, Minseek;Ryu, Hojin;Oh, Min Ji;Im, Ji-Hoon;Lee, Jong-Won;Oh, Youn-Lee
    • Journal of Mushroom
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    • v.20 no.3
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    • pp.178-182
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    • 2022
  • Despite the long history of mushroom use, studies examining the genetic function of mushrooms and the development of new varieties via bio-molecular methods are significantly lacking compared to those examining other organisms. However, owing to recent developments, attempts have been made to use a novel gene-editing technique involving CRISPR/Cas9 technology and genetic scissors in mushroom studies. In particular, research is actively being conducted to utilize ribonucleoprotein particles (RNPs) that can be genetically edited with high efficiency without foreign gene insertion for ease of selection. However, RNPs are too large for Cas9 protein to pass through the cell membrane of the protoplasmic reticulum. Furthermore, guide RNA is unstable and can be easily decomposed, which remarkably affects gene editing efficiency. In this study, nanoparticles were used to mitigate the shortcomings of RNP-based gene editing techniques and to obtain transformants stably. We used Lentinula edodes (shiitake mushroom) Sanjo705-13 monokaryon strain, which has been successfully used in previous genome editing experiments. To identify a suitable osmotic buffer for the isolation of protoplast, 0.6 M and 1.2 M sucrose, mannitol, sorbitol, and KCl were treated, respectively. In addition, with various nanoparticle-forming materials, experiments were conducted to confirm genome editing efficiency via the formation of nanoparticles with calcium phosphate (CaP), which can be bound to Cas9 protein without any additional amino acid modification. RNPs/NP complex was successfully formed and protected nuclease activity with nucleotide sequence specificity.

Biochemical Application of IgG Fc-binding peptide: From Biochip to Targeted Nano Carrier

  • Chung, Sang Jeon
    • Proceedings of the Korean Vacuum Society Conference
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    • 2013.02a
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    • pp.84-84
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    • 2013
  • FcBP consisting of 13 amino acids specifically binds to Immunoglobulin G Fc domain. Initially, we utilized this peptide for preparation of antibody chip as a PEG composite for enhanced solubility. After then, the peptide conjugate was immobilized on agarose resin, resulting in highly efficient affinity column for antibody purification. The efficiency was comparable to commercial Protein A column. Recently, this peptide was conjugated with cell penetratingpeptide (CPP) on a backbone of GFP, affording antibody transducer, which carries antibody into live cells by simple mixing of antibody and the transducer in cell culture media. Antibody transduction into cells was monitored by live cell imaging. More recently, the FcBP was fused to ferritin cage, which consists of 24 ferritin protein molecules. The FcBP-ferritin cage showed greatly increased binding affinity to human IgG. Its binding was analyzed by QCM and SPR analysis. Finally, it was selectively delivered by Herceptin to SKBR3, a breast cancer cell, over MCF10A, non-tumorigenic cells.

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Polyelectrolyte Micropatterning Using Agarose Plane Stamp and a Substrate Having Microscale Features on Its Surface

  • Lee, Min-Jung;Lee, Nae-Yoon;Lee, Sang-Kil;Park, Sung-Su;Kim, Youn-Sang
    • Bulletin of the Korean Chemical Society
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    • v.26 no.10
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    • pp.1539-1542
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    • 2005
  • We have introduced polyelectrolyte micro-patterning technique employing agarose plane stamp and a hard substrate having microscale features on its surface. With this method, chemically micropatterned surfaces with both positive and negative functionalities were successfully embedded in well-defined microstructures, and selective impartment of charge functionalities was confirmed by patterning bead bearing surface charge. Furthermore, this technique allows highly sensitive immobilization of protein onto targeted surface simply by endowing functionalities, which extends the potential of its use as a tool for high-throughput protein microarray and proteomics. Because plane agarose stamp is free of structures on its surface, there is no concern for pattern collapse, and the combination of agarose plane stamp with patterned substrate is more suited for selective protein patterning compared with adopting surface-patterned agarose stamp with flat substrate. Our technique using agarose plane stamp and a substrate having microscale features on its surface suggests a range of possible applications, including the micropatterning of biofunctionalized copolymer having polyelectrolyte block, immobilization of micro- and nanoparticle with biofunctionalities such as biotin and streptavidine, and establishing optoelectronic microstructures with micro-beads on various surfaces.

In Vitro Formation of Protein Nanoparticle Using Recombinant Human Ferritin H and L Chains Produced from E. coli

  • RO HYEON SU;PARK HYUN KYU;KIM MIN GON;CHUNG BONG HYUN
    • Journal of Microbiology and Biotechnology
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    • v.15 no.2
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    • pp.254-258
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    • 2005
  • We have conducted in vitro reconstitution study of ferritin from its subunits FerH and FerL. For the reconstitution, FerH was produced from an expression vector construct in Escherichia coli and was purified from a heat treated cell extract by using one-step column chromatography. FerL was expressed as inclusion bodies. The denatured form of FerL was obtained by a simple washing step of the inclusion bodies with 3 M urea. The reconstitution experiment was conducted with various molar ratios of urea-denatured FerH and FerL to make the ferritin nanoparticle with a controlled composition of FerH and FerL. SDS-PAGE analysis of the reconstituted ferritins revealed that the reconstitution required the presence of more than 40 molar$\%$ of FerH in the reconstitution mixture. The assembly of the subunits into the ferritin nanoparticle was confmned by the presence of spherical particles with diameter of 10 nm by the atomic force microscopic image. Further analysis of the particles by using a transmission electron microscope revealed that the reconstituted particles exhibited different percentages of population with dense iron core. The reconstituted ferritin nanoparticles made with molar ratios of [FerH]/[FerL]=l00/0 and 60/40 showed that 80 to $90\%$ of the particles were apoferritin, devoid of iron core. On the contrary, all the particles formed with [FerH]/[FerL]=85/ 15 were found to contain the iron core. This suggests that although FerH can uptake iron, a minor portion of FerL, not exceeding $40\%$ at most, is required to deposit iron inside the particle.