• 제목/요약/키워드: 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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    • 제41권4호
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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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    • 제32권10호
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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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    • 제35권9호
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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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    • 제2권1호
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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.

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

  • 김민식;류호진;오민지;임지훈;이종원;오연이
    • 한국버섯학회지
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    • 제20권3호
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    • pp.178-182
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    • 2022
  • 버섯의 오랜 역사에도 불구하고 버섯의 유전적 기능과 분자유전학을 응용한 신품종 개발에 대한 연구는 크게 부족한 상황이다. 그러나 최근 유전자 가위인 CRISPR/Cas를 이용한 새로운 유전자 교정 기술이 개발됨에 따라 버섯 연구에서 이 기술을 이용한 다양한 시도가 이루어지고 있다. 특히 선택의 용이성을 위해 외래 유전자 삽입 없이도 고효율로 유전자 편집이 가능한 RNPs를 활용한 연구가 활발히 진행되고 있다. 그러나 RNPs는 원형질체의 세포막을 통과하기에 Cas9이 너무 거대하고 guide RNA가 쉽게 파괴된다는 단점을 가지고 있다. 이러한 단점을 극복하기 위하여 세포막 통과에 용이한 미네랄 성분인 CaP와 PAA를 조합하여 Nanoparticle을 형성함으로써 극복하고자 했다. 표고버섯 단핵 균주인 산조705-13을 이용하여 원형질체 분리에 적합한 Osmotic buffer를 찾기 위하여 0.6M과 1.2M의 Sucrose, Sorbitol, Mannitol, KCl을 처리하였고 그 결과 0.6M Sucrose가 가장 적합한 osmotic buffer임을 확인하였다. 또한 CaP으로 RNPs와 Nanoparticle 복합체를 형성하고 이 복합체가 RNase A로부터 RNPs의 기능을 온전히 보호하는 것을 확인할 수 있었다.

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

  • Chung, Sang Jeon
    • 한국진공학회:학술대회논문집
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    • 한국진공학회 2013년도 제44회 동계 정기학술대회 초록집
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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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    • 제26권10호
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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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    • 제15권2호
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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.