• Title/Summary/Keyword: plasma protein adsorption

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

  • Hyun, J.W.;Kim, N.Y.
    • Journal of the Korean Vacuum Society
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    • v.17 no.6
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    • pp.549-554
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    • 2008
  • Nickel Chloride coated protein chip plates were developed by using a spin coating method after $H_2$ plasma treatment. The adsorption ability of histidine tagged protein was investigated at various times of plasma treatment. The properties of the nickel chloride and protein on the surface of the slides were assayed using particle size analysis and the extent of the protein adsorption was determined by using a bio imaging analyzer system. The results show that the ability of protein adsorption decreased as increasing the time of $H_2$ plasma treatment. The mechanism on the ability of protein adsorption at the plate surface is discussed on results and discussions. The results also suggest that the surface stabilization of protein chip plates treated by plasma technology may be applicable in biosensor markets.

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

  • Insup Noh;Lee, Jin-Hui
    • Macromolecular Research
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    • v.9 no.4
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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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    • v.11 no.6
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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
    • Proceedings of the Korean Vacuum Society Conference
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    • 2016.02a
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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
    • Journal of Biomedical Engineering Research
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    • v.14 no.4
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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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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.

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

  • Lee, Mi-Jung;Hwang, In-Young;Kim, Sung-Kyu;Jung, Suk-Hyun;Jeong, Seo-Young;Seong, Ha-soo;Cho, Sun-Hang;Shin, Byung-Cheol
    • Journal of Pharmaceutical Investigation
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    • v.39 no.6
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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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    • v.6 no.6
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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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    • v.1 no.2
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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.

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

  • 이진호;강길선
    • Journal of Biomedical Engineering Research
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    • v.10 no.1
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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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