• Title/Summary/Keyword: cationic protein

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Surface Properties of Liposomes Modified with Poly(ethylenimine) (폴리에틸렌이민으로 개질된 리포솜의 표면 특성)

  • 박윤정;남다은;서동환;한희동;김태우;김문석;신병철
    • Polymer(Korea)
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    • v.28 no.6
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    • pp.502-508
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    • 2004
  • Cationic liposomes for cancer treatment have been developed in the field of chemotharpy. It was well combined on the surface of anionic tumor cell membrane by electrostatic interaction. Thus, the object of this study was to prepare the cationic liposomes capable of forming an ionic complex with the anionic cell membrane. To prepare the cationic liposomes, 1,2-distearoyl-sn-glycero-3-phosphoethanolamine (DSPE) as a cationic lipid material and polyethylenimine (PEI) as a cationic polymer were synthesized. Ionic property on the surface of liposomes was determined by the zeta potential. The adsorption characteristics of plasma protein for liposome in bovine serum were determined by the particle size and turbidity change. To estimate the stability of liposome in buffered solution, the change of particle size was measured at room temperature for seven days. The cationic liposomes were absorbed a large amount of plasma protein in bovine serum because plasma protein having anionic charge was fixed on the surface of cationic liposomes. This result indicate that the modification on the surface of liposomes using cationic polyethylenimine enhances the protein adsorption in bovine serum. Additionaly, cationic liposomes showed good stability in buffered solution for seven days.

Different Influences of Biotinylation and PEGylation on Cationic and Anionic Proteins for Spheroid Penetration and Intracellular Uptake to Cancer Cells

  • Jung, Won Ho;You, Gayeon;Mok, Hyejung
    • Journal of Microbiology and Biotechnology
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    • v.32 no.9
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    • pp.1209-1216
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    • 2022
  • To better understand the effects of PEGylation and biotinylation on the delivery efficiency of proteins, the cationic protein lysozyme (LZ) and anionic protein bovine serum albumin (BSA) were chemically conjugated with poly(ethylene glycol) (PEG) and biotin-PEG to primary amine groups of proteins using N-hydroxysuccinimide reactions. Four types of protein conjugates were successfully prepared: PEGylated LZ (PEG-LZ), PEGylated BSA (PEG-BSA), biotin-PEG-conjugated LZ (Bio-PEG-LZ), and biotin-PEG-conjugated BSA (Bio-PEG-BSA). PEG-LZ and Bio-PEG-LZ exhibited a lower intracellular uptake than that of LZ in A549 human lung cancer cells (in a two-dimensional culture). However, Bio-PEG-BSA showed significantly improved intracellular delivery as compared to that of PEG-BSA and BSA, probably because of favorable interactions with cells via biotin receptors. For A549/fibroblast coculture spheroids, PEG-LZ and PEG-BSA exhibited significantly decreased tissue penetration as compared with that of unmodified proteins. However, Bio-PEG-BSA showed tissue penetration comparable to that of unmodified BSA. In addition, citraconlyated LZ (Cit-LZ) showed reduced spheroid penetration as compared to that of LZ, probably owing to a decrease in protein charge. Taken together, chemical conjugation of targeting ligands-PEG to anionic proteins could be a promising strategy to improve intracellular delivery and in vivo activity, whereas modifications of cationic proteins should be more delicately designed.

Separation of Lactoferrin from Model Whey Protein Mixture by Reverse Micelles Formed by Cationic Surfactant

  • Noh, Kyung-Hyun;Rhee, Min-Suk;Imm, Jee-Young
    • Food Science and Biotechnology
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    • v.14 no.1
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    • pp.131-136
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    • 2005
  • The selective extraction behavior of lactoferrin (Lf) from whey protein mixture was examined using reverse micelles formed by the cationic surfactant, cetyldimethylammonium bromide (CDAB). The major whey proteins, including ${\beta}$-lactoglobulin, ${\alpha}$-lactalbumin and bovine serum albumin, were solubilized from aqueous phase to organic phase while Lf was recovered in the aqueous phase. The solubilization behaviors of the proteins were manipulated by the process parameters such as the pH and salt concentration of the aqueous phase and the surfactant concentration in the organic phase. Efficient forward extraction was achieved with sodium borate buffer (50 mM, pH 9) containing 50 mM KCl and organic phase containing 100 mM CDAB. Based on SDS-PAGE and densitometry, about 96% of the initial Lf remained in the aqueous phase after forward extraction. The dialyzed Lf fully maintained its bacteriostatic activity against E. coli O157:H7.

Optimization of Gene Transfection Using Fluorescence-Activated Cell Sorter(FACS) Analysis of Green Fluorescent Protein(GFP) (Green Fluorescent Protein(GFP)의 Fluorescence-Activated Cell Sorter(FACS) 분석을 통한 유전자 이입의 최적화)

  • 김태경;박민태;이균민
    • KSBB Journal
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    • v.14 no.3
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    • pp.377-379
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    • 1999
  • In order to improve the transfection efficiency of CHO/dhfr- cells using cationic lipid, optimal concentrations of the cationic lipid($LipofectAmine^{TM}$) and DNA(pEGFP-C1) need to be determined. The use of green fluorescent protein(GFP) gene as a reporter gene facilitated the quantification of transfection efficiency. The green fluorescence intensity of each cell transfected at various lipid-DNA concentrations was measured using fluorescence-activated cell sorter(FACS) analysis. A combination of $2.0{\mu}L$ cationic lipid and 0.4{$\mu}g$ DNA in a well resulted in the highest trasfection efficiency. Taken together, the method using FACS analysis of GFP is simple and fast, facilitating the optimization of transfection.

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Preparation of Protein-coated Cationic Liposomes Containing Doxorubicin and Their Binding Property of Blood Plasma Protein (독소루비신을 함유하고 단백질로 수식된 양이온성 리포솜의 제조 및 혈장 단백흡착 특성)

  • Kim, Sung-Kyu;Jung, Soon-Hwa;Jung, Suk-Hyun;Seong, Ha-Soo;Chi, Sang-Cheol;Cho, Sun-Hang;Shin, Byung-Cheol
    • Journal of the Korean Chemical Society
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    • v.52 no.1
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    • pp.57-65
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    • 2008
  • are nanometer or micrometer scale vesicles that can be used as drug delivery carriers. However, plain liposomes are plagued by rapid opsonization, making their circulation time in bloodstream be shortened. In this study, model protein, bovine serum albumin (BSA)-coated liposomes were prepared by coating cationic liposomes with BSA molecules at higher pH than isoelectric point of BSA. The BSA molecules coated on the liposomal surface were denatured by thermal treatment at above 60oC. While both plain and cationic liposomes had about mean particle diameter of 1041 nm, BSA-coated cationic liposomes (BCL) had mean particle diameter of 1091 nm. Encapsulation of model drug, doxorubicin (DOX), in liposomes were carried out by using remote loading method and the loading efficiency of DOX to liposomes was about 90%. The mean particle diameter of BCL did not increase in blood plasma and adsorption of plasma protein was much less than plain or cationic liposomes. These results suggest that BCL can be used as a long-circulating liposomes in bloodstream.

A Drug Efflux Pump for Cationic Drugs including Disinfectants in Bacillus subtilis (양이온약제내성을 유도하는 Bacillus subtilis의 Drug Efflux Pump)

  • Yong Joon Chung
    • Microbiology and Biotechnology Letters
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    • v.31 no.3
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    • pp.230-234
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    • 2003
  • The Bacillus subtilis YvaE protein, the small multidrug resistance (SMR) family (TC #2.A. 7.1), is shown to catalyze efflux of multiple cationic drugs including many disinfectants, when it was cloned and expressed in Escherichia coli. When the yvaD gene was coexpressed with yvaE gene, the yvaD protein, encoded within a single operon with the yvaE gene, is shown to counteract the action ofYvaE. By ethidium efflux analysis, the cells harvoring a vector with yvaE gene showed a rapid ethidium efflux, compared with the control cells. These results clearly suggest that YvaE mediates drug export from the cell cytoplasm.

Effect of processed foods on serum levels of eosinophil cationic protein among children with atopic dermatitis

  • Lee, Ji-Min;Jin, Hyun-Jung;Noh, Geoun-Woong;Lee, Sang-Sun
    • Nutrition Research and Practice
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    • v.5 no.3
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    • pp.224-229
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    • 2011
  • The prevalence of atopic dermatitis (AD) in school-age children has increased in industrialized countries. As diet is one of the main factors provoking AD, some studies have suggested that food additives in processed foods could function as pseudoallergens, which comprise the non-immunoglobulin E-mediated reaction. Eosinophil cationic protein (ECP) is an eosinophil granule protein released during allergic reactions to food allergens in patients with AD. Thus, serum ECP levels may be a useful indicator of ongoing inflammatory processes in patients with AD. The purpose of this study was to investigate the effect of consuming MSG in processed foods on serum ECP levels among children with AD. This study was performed with 13 patients with AD (age, 7-11 years) who had a normal range of total IgE levels (< 300 IU/ml). All participants ate normal diets during the first week. Then, six patients were allocated to a processed food-restricted group (PRDG) and seven patients were in a general diet group (GDG). During the second week, children in the PRDG and their parents were asked to avoid eating all processed foods. On the third week, children in the PRDG were allowed all foods, as were the children in the GDG throughout the 3-week period. The subjects were asked to complete a dietary record during the trial period. Children with AD who received the dietary restriction showed decreased consumption of MSG and decreased serum ECP levels and an improved SCORing score on the atopic dermatitis index (P < 0.05). No differences in serum ECP levels or MSG consumption were observed in the GDG. Serum total IgE levels were not changed in either group. In conclusion, a reduction in MSG intake by restricting processed food consumption may lead to a decrease in serum ECP levels in children with AD and improve AD symptoms.

Measurement of Molecular Weight and Heating Properties of Korean White Ginseng Protein (백삼 단백질의 가열특성과 분자량 측정)

  • 박상욱
    • The Korean Journal of Food And Nutrition
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    • v.17 no.1
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    • pp.66-71
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    • 2004
  • This study was carried out to elucidate the molecular weight and the heating properties of Korean white ginseng protein by CM-cellulose column chromatography and electrophoresis. Thermostable protein contents were 0.17% in xylem-pith and 0.15% in cortex-epidermis of tap root by 90min of heating. The contents of thermostable protein were decrease after 90min of heating. By Electrophoresis, seven bands of 66, 45, 29, 24, 22, 20, 12kD were observed up to 30min of heating, but the band of 22kD was disappeared after 60min. of heating. The cationic protein content of thermostable protein fraction (28.24%) was higher than the anion protein content(0.80%). The molecular weight of thermostable protein fractions were 66kD, 55kD, 36kD and those of thermolabile protein fractions were 29kD, 24kD, 22kD, 20kD.

The Clinical Study Of Atopic Dermatitis And Eosinophil Cationic Protein in Atopic Dermatitis (아토피피부염 환자 20명에 대한 임상적 고찰과 ECP의 변화)

  • Kwon Mi-Won;Lee Hai-Ja;Park Eun-Jung;Hwang Woo-Jun
    • The Journal of Pediatrics of Korean Medicine
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    • v.14 no.2
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    • pp.121-132
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    • 2000
  • Background : The level of serum eosinophil cationic protein(ECP) is elevated in Atopic Dermatitis patients. Objective : The aim of this study was to investigate the usefulness of serum ECP as a tool of evaluate the efficacy of herb medicine for atopic dermatitis. Material and Method : We investigated 20 patients suffering from atopic dermatitis and analyzed the relationship among the serum level of ECP, IgE, Eosinophil count, and clinical disease activity. Result: Significant elevation in the serum level of ECP, IgE, Eosinophil count is observed in Atopic Dermatitis. Conclusion : The serum level of ECP may be considered to be an useful tool in evaluate effect of herb medicine for atopic dermatitis.

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Recent Advances and Future Strategy in Gene Delivery System (유전자 수송계의 현재까지의 연구동향 및 앞으로의 개발전략)

  • Choi, Woo-Jeong;Kim, Chong-Kook
    • Journal of Pharmaceutical Investigation
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    • v.30 no.1
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    • pp.1-12
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    • 2000
  • Gene therapy is a method for the treatment of diseases with introducing the gene-engineered materials into a patient with gene-deficiency disease (e.g. cystic fibrosis) or cancer to produce a therapeutic protein in a patient's cells. Successful gene therapy requires establishing both gene expression systems and delivery systems. Viral and non-viral vectors have been used for gene delivery. Viral vectors have a high transfection efficiency, but are limited in relations to issues of safety, toxicity and immunogenecity. Non-viral vectors are easy to prepare and relatively safe. However, non-viral vectors have a low transfection efficiency. Cationic liposomes are the most available among non-viral vectors. Cationic liposomes have been used to transfect cells both in vitro and in vivo experiments. Besides, several formulations containing cationic lipid are being used in clinical trials in cases of cystic fibrosis or cancer. A crucial subject to the further development of gene delivery vectors will be a long-term gene expression with following characteristics; protecting and deliverying DNA efficiently, non-toxic and non-immunogenic, and easy to produce in large scale.

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