• 제목/요약/키워드: Protein Adsorption

검색결과 208건 처리시간 0.027초

Fetal Bovine Serum을 포함한 세포 배양액에 담근 Indium Tin Oxide 전극 계면의 전기화학적 특성 (Electrochemical Properties of Indium Tin Oxide Electrodes Immersed in a Cell Culture Medium with Fetal Bovine Serum)

  • 최원석;조성보
    • 대한의용생체공학회:의공학회지
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    • 제34권1호
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    • pp.34-39
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    • 2013
  • For the biocompatibility test of implantable devices or for the sensitivity evaluation of biomedical sensors, it is required to understand the mechanism of the protein adsorption and the interaction between the adsorbed proteins and cells. In this study, the adsorption of proteins in a cell culture medium with fetal bovine serum onto an indium tin-oxide electrode was characterized by using linear sweep voltammetry and impedance spectroscopy. We immersed the fabricated ITO electrodes in the culture medium for 30, 60, or 90 min, and then measured the electrochemical properties of electrodes with 10 mM $Fe(CN){_6}^{3-/4-}$ and 0.1 M KCl electrolyte. With an increase of contacting time, the anodic peak current was decreased and the charge transfer resistance was increased. However, both parameters were recovered to the values before contact with the medium after the treatment of Trypsin/Ethylenediaminetetraacetic acid hydrolyzing proteins.

콘택트렌즈의 물리화학적 특성에 따른 라이소자임과 알부민의 흡착 특성 (Adsorption Properties of the Lysozyme and Albumin with Physicochemical Properties of the Contact Lens)

  • 성유진;유근창;전진
    • 한국안광학회지
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    • 제18권3호
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    • pp.261-270
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    • 2013
  • 목적: FDA 기준에 의해 분류된 시판용 콘택트렌즈와 실험실에서 제조한 콘택트렌즈의 물리화학적 특성에 따라 라이소자임과 알부민의 흡착 특성을 살펴보고자 한다. 방법: 실험실에서 제조한 렌즈는 HEMA(2-hydroxyethyl methacrylate)와 TRIM(3-(trimethoxysilyl) propyl methacrylate) 등의 모노머를 사용하였으며 캐스트몰드 방법으로 제조하였다. 라이소자임과 알부민을 함유한 인공눈물을 제조하였다. 각각의 렌즈에 대해 흡착시간(48시간)과 인공눈물의 pH(pH 6, 6.8, 7.4, 8.2, 9)에 따라 단백질 흡착량 변화를 추적하고, 콘택트렌즈에 흡착된 각각의 단백질은 HPLC로 정량하였다. 결과: 두 단백질의 흡착에 대한 평형상태 도달하는 시간은 하이드로겔 렌즈에 비해 실리콘하이드로겔 렌즈에서 더 오래 걸렸다. 평형상태에서 두 단백질에 대한 흡착량은 실리콘하이드로겔 렌즈에 비해 하이드로겔 렌즈, 비이온성 렌즈에 비해 이온성 렌즈에서 높게 나타났다. 또한, 고함수 렌즈에서는 라이소자임이, 저함수 렌즈에서는 알부민의 흡착량이 많았으며, 이온성 고함수의 Group IV 하이드로겔 렌즈(H4)에서는 라이소자임만이 흡착되었다. 인공눈물의 pH에 따른 두 단백질의 흡착량은 각 단백질의 등전점에 가까워질수록 증가하였다. 결론: 라이소자임의 흡착량은 콘택트렌즈의 함수율보다는 렌즈 표면의 이온성에 더 큰 영향을 받으며, 알부민은 렌즈 표면의 이온성보다 함수율에 더 많은 영향을 받는다. 실리콘하이드로겔 렌즈에서 단백질의 흡착은 콘택트렌즈의 극성뿐만 아니라 실리콘 모노머에 포함된 Si 원자수와 그 화학적 구조에 의해 결정되는 세공의 크기 등이 함께 고려되어야 한다.

나노기공 표면에서의 세포 행동양식에 관한 연구 (Investigation of Cell Behavior on Nanoporous Surface)

  • 정성희;윤원중;민준홍
    • KSBB Journal
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    • 제27권1호
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    • pp.45-50
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    • 2012
  • In this paper, we investigated the effect of nanostructure on the cell behaviors such as adhesion and growth rate. Nanoporous structures with various diameters (30, 40, 45, 50, 60 nm) and 500 nm of the depth were fabricated using the anodizing method. The water contact angle of the surface consisting of nanopores with 30 nm diameter was 40 degree and those were 60~70 degree in cases of nanopores with over 40 nm diameter. Hela cells were cultivated on various nanoporous structure surface to investigate the cell behavior on nanostructure. As a result, Hela cells preferred 30 nm diameter nanoporous surface that has lower water contact angle. This result was confirmed by protein adsorption experiment and scanning electron microscope investigation.

Solid-phase Refolding of Poly-lysine Tagged Fusion Protein of hEGF and Angiogenin

  • Park Sang Joong;Ryu Kang;Suh Chang Woo;Chai Young Gyu;Kwon Oh Byung;Park Seung Kook;Lee Eun Kyu
    • Biotechnology and Bioprocess Engineering:BBE
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    • 제7권1호
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    • pp.1-5
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    • 2002
  • A fusion protein, consisting of a human epidermal growth factor (hEGF) as the recognition domain and human angiogenin as the toxin domain, can be used as a targeted therapeutic against breast cancer cells among others. The fusion protein was expressed as inclusion body in recombinant E. coli, and when the conventional, solution-phase refolding process was used the refolding yield was very low due to severe aggregation. It was probably because of the opposite electric charge at a neutral pH resulting from the vastly different pI values of each domain. The solid-phase refolding process that exploited the ionic interactions between ionic exchanger surface and the fusion protein was tried, but the adsorption yield was also very low, below $ 30\%$, regardless of the resins and pH conditions used. Therefore, to provide a higher ionic affinity toward the solid matrix, six lysine residues were tagged to the N-terminus of the hEGF domain. When heparin-Sepharose was used as the matrix, the adsorption capacity increased 2.5-3 times to about $88\%$. Besides the intrinsic affinity of angiogenin to heparin, the poly-lysine tag provided additional ionic affinity. And the subsequent refolding yield increased nearly 13-fold, from ca. $4.8\%$ in the conventional refolding of the untagged fusion protein to $63.6\%$. The process was highly reproducible. The refolded protein in the column eluate retained RNase bioactivity of angiogenin.

분리 대두단백질의 기름-물 계면흡착 과 유화안정성에 관한 연구 (A Study on the Adsorption at Oil-Water Interface and the Emulsion Stabilizing Properties of Soy Protein Isolate)

  • 김영숙;조형용;조은경;이신영;변유량
    • 한국식품과학회지
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    • 제18권6호
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    • pp.468-474
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    • 1986
  • 대두단백질의 용해도, 소수성, 에멀젼의 유동특성, 물-기름 계면에서의 단백질 흡착 및 계면장력과 유화 특성과의 관계를 연구 검토 하였다. 등전점 부근에서 단백질의 유화능력과 유화안정도가 최소였으며, 단백질 용액의 pH가 이로부터 산성과 알칼리성 쪽으로 갈수록 유화특성이 증가하는 경향으로, 대두단백질의 용해도와 비슷한 경향을 보였다. 에멀젼의 온도가 증가함에 따라 $50^{\circ}C$까지는 유화형성능력이 증가하다가 그 이상에서는 약간 감소하였으며, 유화안 정도는 온도 증가에 따라 계속 감소하는 경향을 보였다. 인산염이 존재할 경우를 제외하고는 전반적으로 염은 유화특성을 감소시켰다. 단백질의 소수성은 온도가 증가함에 따라 증가하였으며, pH가 낮아질수록 약간 증가하는 경향을 보이다가 등전정 이하 산성영역에서 급격히 증가하였다. 기름-물 계면에서 단백질의 최대 흡착량은 pH2.0에서 $0.78mg/m^2$로 가장 높았으며. 다음 pH7이었고, 등전점인 pH4에서 가장 낮았다. 대두단백질이 존재하지 않을때 물-기름 계면장력은 19.76dyne/cm 이었으나 단백질이 존재할 경우 각 조건에 따라 다소 차이는 있으나 11.45-18.08dyne/cm로 전반적으로 저하되었다.

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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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고분자 표면의 혈관내피세포 부착력에 관한 연구 (Evaluation of Endothelial Cell Attachment on Polymer Surface)

  • 최진욱;유규하;민병구
    • 대한의용생체공학회:의공학회지
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    • 제11권1호
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    • pp.131-140
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    • 1990
  • To improve antithrombogenicity of polymer that used in vascular graft and artificial organs, seeding of human endothelial cells on the polyurethane was studied. Human endothelial cells were ismlated from human umbilical veins, using type I collagenase, and identified with goat anti vWF antibodies. Human endothilial cell seeding was tried upon the polyurethane which has good mechanical property and resists stresses. The hydrophobic polyurethane surface was changed hydrophilic by corona discharf:e treatment. Surface hydrophilicity was measured with Wilhemly plate method and the goniometer. To evaluate matrix protein adsorption, fibronectin adsorption test was done. To eveluate cell adhesion, human endothelial cell attachment forces were measured rising a perfusion chamber of , ism diamter. Less cells were detached from the hydrophilically treated polyurethane. This showed that corona discharge on the polyurethane could improve matrix adsorption and endothelial cell attachment.

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A Kinetic Study on the Adsorptionof Compact, Water-soluble Proteins onto Aqueous Surfaces

  • 조태철;Michel A. Cornec
    • Bulletin of the Korean Chemical Society
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    • 제20권9호
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    • pp.999-1004
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    • 1999
  • Two compact sized globular proteins, β-lactoglobulin and α-lactalbumin were kinetically characterized at the aqueous solution surface with the measurement of surface pressure (π) and surface concentration (Γ) via a radiotracer method. The adsorption kinetics was of diffusion control at early times, the rates of increase of πand Γ being lower at longer times due to growing energy barrier. At low concentrations, an apparent time lag was observed in the evolution of π for β-lactoglobulin but not for α-lactalbumin which was shown to be due to the non-linear nature of the p- G relationship for the former. The area per molecule of an adsorbed β-lactoglobulin during adsorption was smaller than that for spread monolayer since β-lactoglobulin was not fully unfolded during the adsorption. For α-lactalbumin, however, no such difference in the molecular areas for adsorbed and spread monolayer was observed indicating thereby that α-lactalbumin unfolded much more rapidly (has looser tertiary structure) than β-lactoglobulin. Surface excess concentrations of α-lactalbumin was found to evolve in two steps possibly due to the change in the orientation of the adsorbed protein from a side-on to an end-on orientation.

실란 카르복실 표면을 사용한 단백질 칩 기판의 기능 분석 (Functional Analysis of Protein Chip Plate Using Silane Carboxylate Surface)

  • 김지현;송예신;윤미영;피재호
    • 한국표면공학회지
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    • 제37권4호
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    • pp.215-219
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    • 2004
  • We fabricated protein chip plates coated with silane carboxylate. The silane compound was immobilized by hydrogen bond and/or other chemical bonds on the surface of the plate. The plates were then prepared by binding $Ni^{2+}$ to surfaces terminated with silane carboxylate groups. The carboxylic acid surface was generated by chemical oxidation of the terminal double-bond functions of the silane-deposited layer. The $Ni^{2+}$ ions on the surface reacted readily to His-tagged proteins. A significant increase in His-tagged protein adsorption was achieved on the surface terminated with silane carboxylate with longer alkyl chain, suggesting better availability of these protein chip plates for proteomic studies.