• 제목/요약/키워드: plasma protein adsorption

검색결과 16건 처리시간 0.024초

단백질 칩 기판의 플라즈마 효과 (Effects of Plasma on the Surface of Protein Chip Plates)

  • 현준원;김나연
    • 한국진공학회지
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    • 제17권6호
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    • pp.549-554
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    • 2008
  • 수소 플라즈마 처리된 유리 기판에 스핀 코팅 시스템을 이용하여 nickel chloride를 코팅하여 단백질칩 플레이트를 제조하였다. 다양한 플라즈마 처리 시간대에서 histidine tagged 단백질의 부착 능력 특성을 연구하였다. 유리 기판 표면에서 nickel chloride와 단백질 특성을 particle size analysis를 이용하여 관찰하였고, 단백질의 부착 능력 정도를 bio imaging analyzer system으로 측정하였다. 실험 결과에 따르면, 플라즈마 처리 시간이 증가할수록 단백질 부착 능력은 감소하는 것으로 나타났다. 기판 표면에서의 단백질 부착능력 특성에 관한 mechanism은 본문의 결과 및 토의에서 논의되었다. 플라즈마 처리된 단백질칩 기판에 대한 표면 안정화는 바이오센서 시장에서 큰 관심을 끌 것으로 기대된다.

Mechanisms of Platelet Adhesion on Elastic Polymer Surfaces: Protein Adsorption and Residence Effects

  • Insup Noh;Lee, Jin-Hui
    • Macromolecular Research
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    • 제9권4호
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    • pp.197-205
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    • 2001
  • Platelet adhesion onto elastic polymeric biomaterials was tested in vitro by perfusing human whole blood at a shear rate of 100 sec$\^$-1/ for possible verification of mechanisms of initial platelet adhesion perfusion of blood on the polymeric substrates was performed after treatments either with or without pre-adsorption of 1% blood plasma, and either with or without residence of the protein-preadsorbed substrate in phosphate buffered solution. The surfaces employed were elastic polymers such as poly(ether urethane urea), poly(ether urethane), silicone urethane copolymer, silicone rubber and poly(ether urethane) with the anti-calcifying agent hydroxyethane bisphosphate. Each polymer surface treated was exposed in vitro to the dynamic, heparinized whole blood perfused for upto 6 min and the surface area of platelets initially adhered was measured by employing in situ epifluorescence video microscopy. The blood perfusion was performed on the surfaces treated at the following three different conditions: directly on the bare surfaces, after protein pre-adsorption and after residence in buffer for 3 days of the surfaces protein pre-adsorbed for 2 h. The effects of blood plasma pre-adsorption on the initial platelet adhesion was surface-dependent. The amount of the adsorbed fibrinogen and the surface coverage area of the adhered platelets were dependent on the surface conditions whether substrates were bare surfaces or protein pre-adsorbed ones. To test an effect of possible morphological (re)orientations of the adsorbed proteins on the initial platelet adhesion, the polymeric substrate pre-adsorbed with 1% blood plasma was immersed in phosphate buffered solution for 3 days and then exposed to physiological blood perfusion. The surface area of the platelets adhered on these surfaces was significantly different from that of the surfaces treated with protein pre-adsorption only. These results indicated that platelet adhesion was dependent on the surface property itself and pre-treatment conditions such as blood perfusion without any pre-adsorption of proteins, and blood perfusion either after protein pre-adsorption or after subsequent substrate residence in buffer of the substrate pre-adsorbed with proteins. Understanding of these results may guide for better designs of blood-contacting materials based on protein behaviors.

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Plasma Protein Adsorption to Anion Substituted Poly(vinyl alcohol) Membranes

  • Ryu, Kyu-Eun;Hyangshuk Rhim;Park, Chong-Won;Chun, Heung-Jae;Hong, Seung-Hwa;Kim, Jae-Jin;Lee, Young-Moo
    • Macromolecular Research
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    • 제11권6호
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    • pp.451-457
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    • 2003
  • Anion-substituted poly(vinyl alcohol) (PVA) membranes, carboxymethylated PVA (C-PVA), and sulfonated PVA (S-PVA) were prepared and the effects of these substitutions on the plasma protein adsorption were studied by one- and two-dimensional gel electrophoresis and immunoblotting. When Cuprophane was used as a negative control, the amount of total proteins bound to samples decreased in the order Cuprophane > PVA > C-PVA > S-PVA, which we attribute to the effects of the surface characteristics of the samples, such as their surface tensions and electrostatic properties, on the adsorption of proteins to the surfaces of the materials. The results revealed that albumin was the most abundant protein in all the samples. The proportion of adsorbed fibrinogen to S-PVA exceeded those of PVA and C-PVA, whereas S-PVA exhibited the lowest IgG adsorption affinity among the samples we studied.

Amine functionalized plasma polymerized PEG film: Elimination of non-specific binding for biosensing

  • Park, Jisoo;Kim, Youngmi;Jung, Donggeun;Kim, Young-Pil;Lee, Tae Geol
    • 한국진공학회:학술대회논문집
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    • 한국진공학회 2016년도 제50회 동계 정기학술대회 초록집
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    • pp.378.2-378.2
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    • 2016
  • Biosensors currently suffer from severe non-specific adsorption of proteins, which causes false positive errors in detection through overestimation of the affinity value. Overcoming this technical issue motivates our research. Polyethylene glycol (PEG) is well known for its ability to reduce the adsorption of biomolecules; hence, it is widely used in various areas of medicine and other biological fields. Likewise, amine functionalized surfaces are widely used for biochemical analysis, drug delivery, medical diagnostics and high throughput screening such as biochips. As a result, many coating techniques have been introduced, one of which is plasma polymerization - a powerful coating method due to its uniformity, homogeneity, mechanical and chemical stability, and excellent adhesion to any substrate. In our previous works, we successfully fabricated plasmapolymerized PEG (PP-PEG) films [1] and amine functionalized films [2] using the plasma enhanced chemical vapor deposition (PECVD) technique. In this research, an amine functionalized PP-PEG film was fabricated by using the plasma co-polymerization technique with PEG 200 and ethylenediamine (EDA) as co-precursors. A biocompatible amine functionalized film was surface characterized by X-ray photoelectron spectroscopy (XPS) and Fourier-transform infrared spectroscopy (FT-IR). The density of the surface amine functional groups was carried out by quantitative analysis using UV-visible spectroscopy. We found through surface plasmon resonance (SPR) analysis that non-specific protein adsorption was drastically reduced on amine functionalized PP-PEG films. Our functionalized PP-PEG films show considerable potential for biotechnological applications such as biosensors.

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Analysis of the Adsorbed Plasma Proteins in the Moving Actuator type Total Artificial Heart

  • Gyu Ha Ryu;Jon
    • 대한의용생체공학회:의공학회지
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    • 제14권4호
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    • pp.307-314
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    • 1993
  • Plasma protein adsorption is the first event in the blood-material interaction and influenc- es subsequent platelet adhesion towards thlㅈombus formation. Thiㅈomboembolic events are strongly influenced by surface characteristics of materials and fluid dynamics inside the blood pump. In vitro flow visualizaion and an amimal experiment with the moving actuator type TAH were Performed in order to investigate fluid dynamic effects on the protein adsorption. The diffel'encl level, j of shear rate inside the ventricle Lvere determined by consid- ering the direction of the major opening of four healt valves in the implanted TAH and the visualized flow patterns as well. Each ventricle of the explanted TAH was sectionalized into 12 segments according to the shear rate level. The adsorbed protein on each segment was quantified using the ELISA method after soaking in 2% (wye)SDS/PBS for two days. Adsorbed protein layer thicknesses Itvere measured by the Immunogotd method under TEM. The SEM observation show that right ventricle (RV) , immobilized with albumin, displayed different degrees of platelet adhesion on each segment, whereas the left ventricle (LV), grafted by PEO-sulronate, indicated nearly , iame platelet adhesion behavior, regardless of shear rates. The surface concentrations of adsorbed proteins in the low shear rate region are hlghel'than those in the high region, which was confirmed statistically. A modified adsorption model of plasma protein onto polyurethane surface was suggested by considering the effect of the fluid dynamic characteristics.

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

  • 박윤정;남다은;서동환;한희동;김태우;김문석;신병철
    • 폴리머
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    • 제28권6호
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    • pp.502-508
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    • 2004
  • 암을 치료하기 위한 양이온성 리포솜은 화학치료요법의 분야에서 발전되어 왔다. 양이온성 리포솜은 음이온성의 세포 표면에 정전기적 상호작용에 의해 결합이 된다. 본 연구의 목적은 음이온성 세포와 이온결합을 할 수 있는 양이온성 리포솜을 제조하는 것이다. 양이온성 리포솜은 지질인 1,2-디스테로일-sn-글리콜-3-포스포에탄올아민 (DSPE)과 양이온성 고분자인 폴리에틸렌이민 (PEI)을 그라프트 중합으로 합성된 물질로부터 제조하였다. 리포솜 표면의 이온 특성은 제타 포텐셜을 측정하여 확인하였다. 소 혈청 수용액에서 리포솜의 혈장 단백질 흡착 특성은 입자 크기와 탁도 변화를 측정하여 확인하였다. 완충 용액 속에서 리포솜 안정성을 예측하기 위하여 입자 크기를 7일 동안 상온에서 측정하였다. 양이온성 리포솜은 소 혈청 수용액에서 많은 양의 혈장 단백질이 흡착되었다. 이는 혈장 단백은 음전하를 가지고 있어서 양이온성 리포솜의 표면과 잘 붙기 때문이다. 양이온성 폴리에틸렌이민을 이용한 리포솜의 표면 변형은 소 혈청 수용액 내에서 단백질 흡착을 강화시킨다는 것을 예상하게 한다. 또한, 제조된 리포솜의 완충 용액 내에서 7일 동안 안정한 상태임을 관찰하였다.

열변성 단백질이 결합된 음이온성 리포솜의 혈장 내 안정성 및 세포 내 이입 평가 (Stability in Plasma and Intracellular Uptake of Thermally Denatured Protein-coated anionic Liposomes)

  • 이미정;황인영;김성규;정석현;정서영;성하수;조선행;신병철
    • Journal of Pharmaceutical Investigation
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    • 제39권6호
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    • pp.423-429
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    • 2009
  • Liposomes have been used as one of the efficient carriers for drug delivery. In this study, anionic liposomes of which surface was modified by using both electrostaic interaction between anionic liposomes and cationically charged BSA molecules at lower pH than isoelectric point (pI) of BSA and denaturation of the BSA-coated liposomes by thermal treatment. The thermally denatured BSA-coated liposomes (DBAL) had mean particle diameter of 125.2${\pm}$1.7 nm and zeta potential value of -22.4${\pm}$4.5 mV. Loading efficiency of model drug, doxorubicin (DOX), into liposomes was 83.0${\pm}$2.6%. Results of in vitro stability study of DBAL in blood plasma showed that the mean particle diameter of DBAL 400 did not increase in blood plasma and adsorption of plasma protein was much less than plain or anionic liposomes. Intracellular uptake of DBAL 400 evaluated by confocal microscopy observation was higher than that of PEG liposomes.

Methacrylamidohistidine in Affinity Ligands for Immobilized Metal-ion Affinity Chromatography of Human Serum Albumin

  • Odaba, Mehmet;Garipacan, Bora;Dede, Semir;Denizli, Adil
    • Biotechnology and Bioprocess Engineering:BBE
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    • 제6권6호
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    • pp.402-409
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    • 2001
  • Different bioligands carrying synthetic adsorbents have been reported in the literature for protein separation, We have developed a novel and new approach to obtain high protein ad-sorption capacity utilizing 2-methacrylamidohistidine(MAH) as a bioligand. MAH was synthe-sized by reacting methacrylocholride and histidine, Spherical beads with an average size of 150-200㎛ were obtained by the radical suspension polymerization of MAH and 2-hydrosyethyl-methacrylate(HEMA) conducted in an aqueous dispersion medium. p(HEMA-co-MAH) beads had a specific surface area of 17.6㎡/g . Synthesized MAH monomer was characterized by NMR. p(HEMA-co-MAH) beads were characterized by swelling test, FTIR and elemental analysis. Then Cu(II) ions were incorporated onto the beads and Cu(II) loading was found to be 0.96 mmol/g.These affinity beads with a swelling ration of 65% and containing, 1.6 mmol MAH/g were used in the adsorption/desorption of human serum albumin(HSA) from both aqueous solutions and hu-man serum. The adsorption of HSA onto p(HEM-co-MAH) was low(8.8 mg/g). Cu(II) chelation onto the beads significantly increased the HSA adsorption (56.3 mg/g). The maximum HSA ad-sorption ws observed at pH 8.0 Higher HSA adsorption was observed from human plasma(94.6 mgHSA/g) Adsorption of other serum proteins were obtained as 3.7 mg/g for fibrinogen and 8.5mg/g for γ-globulin. The total protein adsorption was determined as 107.1mg/g. Desorption of HSA was obtained using 0.1 M Tris/HCl buffer containing 0.5 M NaSCN, High desorption rations(up to 98% of the adsorbed HSA) were observed. It was possible to reuse Cu(II) chelated-p(HEMA-co-MAH) beads without significant decreases in the adsorption capacities.

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Surface modification of polymeric membranes for low protein binding

  • Higuchi, Akon;Tamai, Miho;Tagawa, Yoh-Ichi;Chang, Yung;Ling, Qing-Dong
    • Membrane and Water Treatment
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    • 제1권2호
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    • pp.103-120
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    • 2010
  • Surface modification of microfiltration and ultrafiltration membranes has been widely used to improve the protein adsorption resistance and permeation properties of hydrophobic membranes. Several surface modification methods for converting conventional membranes into low-protein-binding membranes are reviewed. They are categorized as either physical modification or chemical modification of the membrane surface. Physical modification of the membrane surface can be achieved by coating it with hydrophilic polymers, hydrophilic-hydrophobic copolymers, surfactants or proteins. Another method of physical modification is plasma treatment with gases. A hydrophilic membrane surface can be also generated during phase-inverted micro-separation during membrane formation, by blending hydrophilic or hydrophilic-hydrophobic polymers with a hydrophobic base membrane polymer. The most widely used method of chemical modification is surface grafting of a hydrophilic polymer by UV polymerization because it is the easiest method; the membranes are dipped into monomers with and without photo-initiators, then irradiated with UV. Plasma-induced polymerization of hydrophilic monomers on the surface is another popular method, and surface chemical reactions have also been developed by several researchers. Several important examples of physical and chemical modifications of membrane surfaces for low-protein-binding are summarized in this article.

세포적합성 고분자 표면에 관한 연구 I. 고분자 표면 개질과 ESCA 분석 (Polymer Surfaces for Cell Adhesion I. Surface Modification of Polymers and ESCA Analysis)

  • 이진호;강길선
    • 대한의용생체공학회:의공학회지
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    • 제10권1호
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    • pp.43-52
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    • 1989
  • We modified polymer surfaces, polyethylene, polystyrene and polyester, to improve cellcompatibility. For surface modification of the polymers, we used various surface treatment methods; physicochemical oxidation methods such as plasma discharge, corona discharge, sulfuric acid and chloric acid treatments, and biological methods such as adsorption of plasma protein and fibronectin onto the polymer surfaces. The treated polymer surfaces were characterized by electron spectroscopy for chemical analysis ( ESCA ). The physicochemically treated polymers showed different surface chemical structures depending on the treated methods. The sulfuric acid-treated surfaces showed greater carboxyl groups than those of plasma- or corona- treated surfaces, while the chloric acid-treated one showed high density of hydroxyl group on the surface. By the biological treatments, the surfaces were uniformly coated with proteins. The fibronectin adsorbed on the surface seems to have unique properties for cell binding.

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