• Title/Summary/Keyword: non-specific protein adsorption

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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
    • 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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Conjugation of mono-sulfobetaine to alkyne-PPX films via click reaction to reduce cell adhesion

  • Chien, Hsiu-Wen;Keng, Ming-Chun;Chen, Hsien-Yeh;Huang, Sheng-Tung;Tsai, Wei-Bor
    • Biomaterials and Biomechanics in Bioengineering
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    • v.3 no.1
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    • pp.59-69
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    • 2016
  • A surface resisting protein adsorption and cell adhesion is highly desirable for many biomedical applications such as diagnostic devices, biosensors and blood-contacting devices. In this study, a surface conjugated with sulfobetaine molecules was fabricated via the click reaction for the anti-fouling purpose. An alkyne-containing substrate (Alkyne-PPX) was generated by chemical vapor deposition of 4-ethynyl-[2,2]paracyclophane. Azide-ended mono-sulfobetaine molecules were synthesized and then conjugated on Alkyne-PPX via the click reaction. The protein adsorption from 10% serum was reduced by 57%, while the attachment of L929 cells was reduced by 83% onto the sulfobetaine-PPX surface compared to the protein adsorption and cell adhesion on Alkyne-PPX. In conclusion, we demonstrate that conjugation of mono-sulfobetaine molecules via the click chemistry is an effective way for reduction of non-specific protein adsorption and cell attachment.

Single-Protein Molecular Interactions on Polymer-Modified Glass Substrates for Nanoarray Chip Application Using Dual-Color TIRFM

  • Kim, Dae-Kwang;Lee, Hee-Gu;Jung, Hyung-Il;Kang, Seong-Ho
    • Bulletin of the Korean Chemical Society
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    • v.28 no.5
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    • pp.783-790
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    • 2007
  • The immobilization of proteins and their molecular interactions on various polymer-modified glass substrates [i.e. 3-aminopropyltriethoxysilane (APTS), 3-glycidoxypropyltrimethoxysilane (GPTS), poly (ethylene glycol) diacrylate (PEG-DA), chitosan (CHI), glutaraldehyde (GA), 3-(trichlorosilyl)propyl methacrylate (TPM), 3'-mercaptopropyltrimethoxysilane (MPTMS), glycidyl methacrylate (GMA) and poly-l-lysine (PL).] for potential applications in a nanoarray protein chip at the single-molecule level was evaluated using prismtype dual-color total internal reflection fluorescence microscopy (dual-color TIRFM). A dual-color TIRF microscope, which contained two individual laser beams and a single high-sensitivity camera, was used for the rapid and simultaneous dual-color detection of the interactions and colocalization of different proteins labeled with different fluorescent dyes such as Alexa Fluor® 488, Qdot® 525 and Alexa Fluor® 633. Most of the polymer-modified glass substrates showed good stability and a relative high signal-to-noise (S/N) ratio over a 40-day period after making the substrates. The GPTS/CHI/GA-modified glass substrate showed a 13.5-56.3% higher relative S/N ratio than the other substrates. 1% Top-Block in 10 mM phosphate buffered saline (pH 7.4) showed a 99.2% increase in the blocking effect of non-specific adsorption. These results show that dual-color TIRFM is a powerful methodology for detecting proteins at the single-molecule level with potential applications in nanoarray chips or nano-biosensors.

Surface Modification of PDMS for Hydrophilic and Antifouling Surface Using PEO-PPO-PEO Block Copolymer (PEO-PPO-PEO 블록 공중합체를 이용한 PDMS의 친수성 표면 개질 방법)

  • Lee, Byungjin;Jin, Si Hyung;Jeong, Seong-Geun;Kang, Kyoung-Ku;Lee, Chang-Soo
    • Korean Chemical Engineering Research
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    • v.55 no.6
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    • pp.791-797
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    • 2017
  • In this study, we optimized a method of PEO-PPO-PEO block copolymer embedding, for solving non-specific protein and biomolecular adsorption and high hydrophobicic surface property, which is widely known as problems of poly (dimethylsiloxane) (PDMS) that has frequently been used in basic biological and its applied research. We assessed its surface modification by controlling the concentration of embedded block copolymer, water-soaking time, and recovery time as variables by contact angle measurements. In order to evaluate its antifouling ability, adsorption of FITC-BSA molecules was quantified. Furthermore, we generated oil-in-water (O/W) emulsion as a proof-of-concept experiment to confirm that the optimized surface modification works properly.

Synthesis and Polymerization of Methacryloyl-PEG-Sulfonic Acid as a Functional Macromer for Biocompatible Polymeric Surfaces

  • Kim, Jun-Guk;Sim, Sang-Jun;Kim, Ji-Heung;Kim, Soo-Hyun;Kim, Young-Ha
    • Macromolecular Research
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    • v.12 no.4
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    • pp.379-383
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    • 2004
  • Poly(ethylene glycol)s (PEGs) are unique in their material properties, such as biocompatibility, non-toxicity, and water-solublizing ability, which are extremely useful for a variety of biomedical applications. In addition, a variety of functional PEGs with specific functionality at one or both chain ends have been synthesized for many specialized applications. Surface modifications using PEG have been demonstrated to decrease protein adsorption and platelet or cell adhesion on biomaterials. Furthermore, PEGs having anionic sulfonate terminal units have been proven to enhance the blood compatibility of materials, which has been demonstrated by the negative cilia concept. The preparation of telechelic PEGs having a sulfonic acid group at one end and a polymerizable methacryloyl group at the other is an interesting undertaking for providing macromers that can be used in various vinyl copolymerization and gel systems. In this paper, preliminary results on the synthesis and polymerization behavior of a novel PEG macromer is described with the aim of identifying a biocompatible material for applications in various blood-contacting devices.

Development of Voltammetric Nanobio-incorporated Analytical Method for Protein Biomarker Specific to Early Diagnosis of Lung Cancer (폐암 조기 진단을 위한 단백질 바이오마커 측정용 전압-전류법 기반의 나노바이오 분석법 개발)

  • Li, Jingjing;Si, Yunpei;Nde, Dieudonne Tanue;Lee, Hye Jin
    • Applied Chemistry for Engineering
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    • v.32 no.4
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    • pp.461-466
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    • 2021
  • In this article, a portable and cost-effective voltammetric biosensor with nanoparticles was developed for the measurements of heterogeneous nuclear ribonucleoprotein A1 protein (hnRNP A1) biomarker which can potentially be used for lung cancer diagnosis. Gold nanoparticles were first electrodeposited onto screen printed carbon electrode (SPCE) followed by immobilizing a single stranded DNA aptamer specific to hnRNP A1 onto the electrode surface. Ethanolamine was also used when immobilizing DNA aptamer on the surface to prevent signals from non-specific adsorption events. Sequential injection of hnRNP A1 biomarker and anti-hnRNP A1 conjugated with alkaline phosphatase (ALP) onto the aptamer chip surface allows to form the sandwich complex of DNA aptamer/hnRNP A1/ALP-anti-hnRNP A1 on the electrode surface which further reacted with 4-aminophenyl phosphate (APP). The electrocatalytic reaction of the enzyme, ALP, and the substrate, APP, resulting in the oxidative current response changes at -0.05 and -0.17 V (vs. Ag/AgCl) against the hnRNP A1 concentration was measured using cyclic and differential pulse voltammetry, respectively. The Au nanoparticles-integrated voltammetric biosensor was applied to analyze human normal serum solutions possibly suggesting potential applicability for lung cancer diagnosis.

Interaction of Schwann Cells with Various Protein- or Polypeptide-Coated PLGA Surfaces (다양한 단백질과 폴리펩타이드로 코팅된 PLGA 표면과 슈반세포와의 상호관계)

  • Park Ki-Suk;Kim Su-Mi;Kim Moon-Suk;Lee Il-Woo;Rhee John-M.;Lee Hai-Bang;Khang Gil-Son
    • Polymer(Korea)
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    • v.30 no.5
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    • pp.445-452
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    • 2006
  • In this study, we investigated interaction of Schwann cells (SCs) with various cell-adhesive coated polymer surface. We used cell-adhesives that like a fibronectin (FN), fibrinogen(FG), laminin(LM), vitronectin (VN), poly-D-Iysine (PDL), and poly-L-Iysine (PLL) to coat PLGA film surface and evaluated the surface property of coated or not PLGA films by measurement of water contact angle and ESCA. SCs were cultured on coated or non-coated PLGA film surface, and then examined the cell adhesion and proliferation by cell count and SEM observation. Cell count results revealed initial cell adhesion related to protein adsorption on PLGA surface. In addition, serum content in media related to cell proliferation rate. In this result, we recognized that adhesion and proliferation of SCs were affected by specific cell-adhesives. In these results, we recognized that is important to provide the suitable surface environment according to cell types and culture condition for improvement of cell adhesion and proliferation.