• Title/Summary/Keyword: Protein nanoparticle

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Recombinant Human L-ferritin from Saccharomyces cerevisiae: Molecular Characterization and Synthesis of Iron Oxide Nanoparticles (효모에서 생산한 재조합 human L-ferritin의 생화학적 특성 및 나노입자의 철산화물 합성)

  • Kim, Kyung-Suk
    • KSBB Journal
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    • v.26 no.2
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    • pp.119-125
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    • 2011
  • In the synthesis of nanoparticles, much attention has been paid to regulating the particle size. There has been a possible evident that using the central cavity (core) of the protein ferritin has a greatly significant influence on it because the core can generate the nanometer-sized mineral particles of variable metal ions. In this report, recombinant human L-ferritins produced from Saccharomyces cerevisiae were purified and their molecular properties were characterized. The cDNA for human ferritin L chain was also expressed in another host such as Escherichia coli, and the properties of recombinant L-ferritins were compared. From isoelectric focusing experiment, the L-ferritin from the recombinant yeast showed no indication of N-glycosylation. Some post-translational modifications other than N-glycosylation were speculated in the L-ferritins from yeast. A difference was made in the L-ferritins in their iron uptake rates and the initial rate of the L-ferritin from yeast was slightly increased. The reconstitution yield and size distribution of the core minerals were analyzed in the L-ferritins by transmission electron microscopy. The L-ferritin from yeast with higher reconstitution yield (54.5%) showed slightly larger sizes (mean 6.92 nm) with narrower size distribution than the L-ferritin from E. coli. It is, in conclusion, speculated that L-ferritin from yeast is relatively superior to the other, in view of the size of nanoparticle and its relative homogeneity.

A Simple and Rapid Methicillin-Resistant Staphylococcus aureus (MRSA) Screening Test Using a Mannose-Binding Lectin (MBL)-Conjugated Gold Nanoparticle Probe

  • So Yeon Yi;Jinyoung Jeong;Wang Sik Lee;Jungsun Kwon;Kyungah Yoon;Kyoungsook Park
    • Journal of Microbiology and Biotechnology
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    • v.33 no.5
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    • pp.698-705
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    • 2023
  • Rapid diagnosis of methicillin-resistant Staphylococcus aureus (MRSA) is essential for guiding clinical treatment and preventing the spread of MRSA infections. Herein, we present a simple and rapid MRSA screening test based on the aggregation effect of mannose-binding lectin (MBL)-conjugated gold nanoparticles (AuNP), called the MRSA probe. Recombinant MBL protein is a member of the lectin family and part of the innate immune system. It can recognize wall teichoic acid (WTA) on the membrane of MRSA more specifically than that of methicillin-sensitive Staphylococcus aureus (MSSA) under optimized salt conditions. Thus, the MRSA probe can selectively bind to MRSA, and the aggregation of the probes on the surface of the target bacteria can be detected and analyzed by the naked eye within 5 min. To demonstrate the suitability of the method for real-world application, we tested 40 clinical S. aureus isolates (including 20 MRSA specimens) and recorded a sensitivity of 100%. In conclusion, the MRSA probe-based screening test with its excellent sensitivity has the potential for successful application in the microbiology laboratory.

Resveratrol-loaded Nanoparticles Induce Antioxidant Activity against Oxidative Stress

  • Kim, Jae-Hwan;Park, Eun-Young;Ha, Ho-Kyung;Jo, Chan-Mi;Lee, Won-Jae;Lee, Sung Sill;Kim, Jin Wook
    • Asian-Australasian Journal of Animal Sciences
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    • v.29 no.2
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    • pp.288-298
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    • 2016
  • Resveratrol acts as a free radical scavenger and a potent antioxidant in the inhibition of numerous reactive oxygen species (ROS). The function of resveratrol and resveratrol-loaded nanoparticles in protecting human lung cancer cells (A549) against hydrogen peroxide was investigated in this study. The 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid (ABTS) assay was performed to evaluate the antioxidant properties. Resveratrol had substantially high antioxidant capacity (trolox equivalent antioxidant capacity value) compared to trolox and vitamin E since the concentration of resveratrol was more than $50{\mu}M$. Nanoparticles prepared from ${\beta}$-lactoglobulin (${\beta}$-lg) were successfully developed. The ${\beta}$-lg nanoparticle showed 60 to 146 nm diameter in size with negatively charged surface. Non-cytotoxicity was observed in Caco-2 cells treated with ${\beta}$-lg nanoparticles. Fluorescein isothiocynate-conjugated ${\beta}$-lg nanoparticles were identified into the cell membrane of Caco-2 cells, indicating that nanoparticles can be used as a delivery system. Hydrogen peroxide caused accumulation of ROS in a dose- and time-dependent manner. Resveratrol-loaded nanoparticles restored $H_2O_2$-induced ROS levels by induction of cellular uptake of resveratrol in A549 cells. Furthermore, resveratrol activated nuclear factor erythroid 2-related factor 2-Kelch ECH associating protein 1 (Nrf2-Keap1) signaling in A549 cells, thereby accumulation of Nrf2 abundance, as demonstrated by western blotting approach. Overall, these results may have implications for improvement of oxidative stress in treatment with nanoparticles as a biodegradable and non-toxic delivery carrier of bioactive compounds.

Preparation of Folic Acid-loaded WPI (Whey Protein Isolate) Nanoparticles by Cold-induced Gelation (냉각유도젤화에 의한 엽산 함유 분리유청단백 나노담체의 제조)

  • Kim, Bum-Keun;Lee, Won-Jae;Oh, Se-Jong;Kim, Jin-Man;Park, Dong-June
    • Food Science of Animal Resources
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    • v.30 no.1
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    • pp.95-101
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    • 2010
  • Folate loaded WPI (whey protein isolate) nanoparticles were prepared using the cold-induced gelation process. The aim of this work was to investigate the effects of process parameters, such as the concentration of the WPI solution, pH, temperature, etc, on the properties of nanoparticles. The results show that the smallest nanoparticles were obtained when a WPI concentration of 1% was used at a pH of 8.0 (<330 nm). In the case of the concentration of $CaCO_3$, the smallest particles were obtained at a concentration of 5 mM. Alginate produced the smallest mean size with the narrowest particle size distribution, while the largest particles were prepared with k-carrageenan. As the w:o ratio increased, the mean particle size also increased. When the release profile was analyzed, the particles were shown to be stable for more than 6 h at a pH of 1.2, where almost all of the folic acid was released within 2 h in the dissolution media of PBS at a pH of 7.4. Thus, the process parameters appear to be important factors that affect the properties of nanoparticles.

Development of Chitosan Coated Solid Lipid Nano-particles Containing 7-Dehydrocholesterol (7-디하이드로콜레스테롤을 함유한 키토산 코팅 처리 Solid Lipid Nano-particle의 개발에 관한 연구)

  • Lee Geun-Soo;Kim Tae-Hoon;Lee Chun-Il;Pyo Hyeong-Bae;Choe Tae-Boo
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.31 no.2 s.51
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    • pp.141-146
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    • 2005
  • Unstable cosmetic active ingredients could rapidly break down in chemical and photochemical process. Therefore, it has become a very important issue to encapsulate active ingredient for the stabilization. 7-Dehydrocholesterol (7-DHC), a precursor of vitamin $D_3$, has been shown to increase levels of protein and mRNA for heat shock protein in normal human epidermal keratinocytes. However, topical dermal application of 7-DHC is restricted due to its poor solubility and chemical unstability. In this study, 7-DHC was incorporated into nano-emulsion (NE), solid lipid nano-particle (SLN), and chitosan coated solid lipid nano-particle (CASLN), respectively. In order to prepare NE and SLN dispersion, high-pressure homogenization at temperature above the melting point of lipid was used Hydrogenated lecithin and polysorbate 60 were used as stabilizer for NE and SLN. CASLN was prepared by high speed homogenizing after adding chitosan solution to the SLN dispersion and showed positively charged particle properties. Decomposition rate of 7-DHC in NE, SLN and CASLN was studied as a function of time at different temperature. Differential scanning calorimetry (DSC) and X-ray diffraction (XRD) studies were performed to characterize state of lipid modification. It appeared that CASLN is the most effective to stabilize 7-DHC and may be used for a useful topical dermal delivery system.

Formulation and Characterization of Antigen-loaded PLGA Nanoparticles for Efficient Cross-priming of the Antigen

  • Lee, Young-Ran;Lee, Young-Hee;Im, Sun-A;Kim, Kyung-Jae;Lee, Chong-Kil
    • IMMUNE NETWORK
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    • v.11 no.3
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    • pp.163-168
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    • 2011
  • Background: Nanoparticles (NPs) prepared from biodegradable polymers, such as poly (D,L-lactic acid-co-glycolic acid) (PLGA), have been studied as vehicles for the delivery of antigens to phagocytes. This paper describes the preparation of antigen-loaded PLGA-NPs for efficient cross-priming. Methods: NPs containing a similar amount of ovalbumin (OVA) but different sizes were produced using a micromixer-based W/O/W solvent evaporation procedure, and the efficiency of the NPs to induce the cross-presentation of OVA peptides were examined in dendritic cells (DCs). Cellular uptake and biodistribution studies were performed using fluorescein isothiocyanate (FITC)-loaded NPs in mice. Results: The NPs in the range of $1.1{\sim}1.4{\mu}m$ in size were the most and almost equally efficient in inducing the cross-presentation of OVA peptides via $H-2K^b$ molecules. Cellular uptake and biodistribution studies showed that opsonization of the NPs with mouse IgG greatly increased the percentage of FITC-positive cells in the spleen and lymph nodes. The major cell type of FITC-positive cells in the spleen was macrophages, whereas that of lymph nodes was DCs. Conclusion: These results show that IgG-opsonized PLGA-NPs with a mean size of $1.1{\mu}m$ would be the choice of biodegradable carriers for the targeted-delivery of protein antigens for cross-priming in vivo.

Generalized Two-dimensional (2D) Correlation Spectroscopy: Principle and Its Applications (일반화된 이차원 상관 분광학: 원리 및 응용)

  • Young Mee Jung;Seung Bin Kim
    • Journal of the Korean Chemical Society
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    • v.47 no.5
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    • pp.447-459
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    • 2003
  • Generalized 2D correlation spectroscopy has been applied extensively to the analysis of spectral data sets obtained during the observation of a system under some external perturbation. It is used in various fields of spectroscopy including IR, Raman, UV, fluorescence, X-ray diffraction, and X-ray absorption spectroscopy (XAS) as well as chromatography. 2D hetero-spectral correlation analysis compares two completely different types of spectra obtained for a system under the same perturbation. Because of the wide range of applications of this technique, it has become one of the standard analytical techniques for the analytical chemistry, physical chemistry, biochemistry, and so on, and for studies of polymers, biomolecules, nanomaterials, etc. In this paper, we will introduce the principle of generalized 2D correlation spectroscopy and its applications that we have studied.

Dual Role of Acidic Diacetate Sophorolipid as Biostabilizer for ZnO Nanoparticle Synthesis and Biofunctionalizing Agent Against Salmonella enterica and Candida albicans

  • Basak, Geetanjali;Das, Devlina;Das, Nilanjana
    • Journal of Microbiology and Biotechnology
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    • v.24 no.1
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    • pp.87-96
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    • 2014
  • In the present study, a yeast species isolated from CETP, Vellore, Tamilnadu was identified as Cryptococcus sp. VITGBN2 based on molecular techniques and was found to be a potent producer of acidic diacetate sophorolipid in mineral salt media containing vegetable oil as additional carbon source. The chemical structure of the purified biosurfactant was identified as acidic diacetate sophorolipid through GC-MS analysis. This sophorolipid was used as a stabilizer for synthesis of zinc oxide nanoparticles (ZON). The formation of biofunctionalized ZON was characterized using UV-visible spectroscopy, XRD, scanning electron microscopy (SEM), and Fourier transform infrared spectroscopy. The antimicrobial activities of naked ZON and sophorolipid functionalized ZON were tested based on the diameter of inhibition zone in agar well diffusion assay, microbial growth rate determination, protein leakage analysis, and lactate dehydrogenase assay. Bacterial pathogen Salmonella enterica and fungal pathogen Candida albicans showed more sensitivity to sophorolipid biofunctionalized ZON compared with naked ZON. Among the two pathogens, S. enterica showed higher sensitivity towards sophorolipid biofunctionalized ZON. SEM analysis showed that cell damage occurred through cell elongation in the case of S. enterica, whereas cell rupture was found to occur predominantly in the case of C. albicans. This is the first report on the dual role of yeast-mediated sophorolipid used as a biostabilizer for ZON synthesis as well as a novel functionalizing agent showing antimicrobial property.

Organic Memory Device Using Self-Assembled Monolayer of Nanoparticles (나노입자 자기조립 단일층을 이용한 유기메모리 소자)

  • Jung, Hunsang;Oh, Sewook;Kim, Yejin;Kim, Minkeun;Lee, Hyun Ho
    • Applied Chemistry for Engineering
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    • v.23 no.6
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    • pp.515-520
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    • 2012
  • In this review, the fabrication of silicon based memory capacitor and organic memory thin film transistors (TFTs) was discussed for their potential identification tag applications and biosensor applications. Metal or non-metal nanoparticles (NPs) could be capped with chemicals or biomolecules such as protein and oligo-DNA, and also be self-assembly monolayered on corresponding target biomolecules conjugated dielectric layers. The monolayered NPs were formed to be charging elements of a nano floating gate layer as forming organic memody deivces. In particular, the strong and selective binding events of the NPs through biomolecular interactions exhibited effective electrostatic phenomena in memory capacitors and TFTs formats. In addition, memory devices fabricated as organic thin film transistors (OTFTs) have been intensively introduced to facilitate organic electronics era on flexible substrates. The memory OTFTs could be applicable eventually to the development of new conceptual devices.

Intranasal Immunization With Nanoparticles Containing an Orientia tsutsugamushi Protein Vaccine Candidate and a Polysorbitol Transporter Adjuvant Enhances Both Humoral and Cellular Immune Responses

  • Cheol Gyun Kim;Won Kyong Kim;Narae Kim;Young Jin Pyung;Da-Jeong Park;Jeong-Cheol Lee;Chong-Su Cho;Hyuk Chu;Cheol-Heui Yun
    • IMMUNE NETWORK
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    • v.23 no.6
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    • pp.47.1-47.16
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    • 2023
  • Scrub typhus, a mite-borne infectious disease, is caused by Orientia tsutsugamushi. Despite many attempts to develop a protective strategy, an effective preventive vaccine has not been developed. The identification of appropriate Ags that cover diverse antigenic strains and provide long-lasting immunity is a fundamental challenge in the development of a scrub typhus vaccine. We investigated whether this limitation could be overcome by harnessing the nanoparticle-forming polysorbitol transporter (PST) for an O. tsutsugamushi vaccine strategy. Two target proteins, 56-kDa type-specific Ag (TSA56) and surface cell Ag A (ScaA) were used as vaccine candidates. PST formed stable nano-size complexes with TSA56 (TSA56-PST) and ScaA (ScaA-PST); neither exhibited cytotoxicity. The formation of Ag-specific IgG2a, IgG2b, and IgA in mice was enhanced by intranasal vaccination with TSA56-PST or ScaA-PST. The vaccines containing PST induced Ag-specific proliferation of CD8+ and CD4+ T cells. Furthermore, the vaccines containing PST improved the mouse survival against O. tsutsugamushi infection. Collectively, the present study indicated that PST could enhance both Ag-specific humoral immunity and T cell response, which are essential to effectively confer protective immunity against O. tsutsugamushi infection. These findings suggest that PST has potential for use in an intranasal vaccination strategy.