• Title/Summary/Keyword: cross linker

Search Result 92, Processing Time 0.029 seconds

Detection of Carp Vitellogenin with Piezoelectric Immunosensor (압전류적 면역센서에 의한 잉어 Vitellogenin 검출)

  • Kim, Nam-Soo;Park, In-Seon;Kim, Woo-Yeon
    • Applied Biological Chemistry
    • /
    • v.49 no.3
    • /
    • pp.254-258
    • /
    • 2006
  • Vitellogenin has been known as a potent biomarker protein for the estrogenic activity in fish exposed to endocrine disruptors. In this study, a piezoelectric immunosensor making use of an anticarp vitellogenin antibody and an AT-cut quartz crystal microbalance as the biological component and transducer was prepared, followed by its application to the analysis of carp vitellogenin as follows. Antibody immobilization was conducted by chemisorption of a thiolated antibody with a heterobifunctional thiolation cross-linker, sulfosuccinimidyl 6-[3-(2-pyridyldithio)propionamido]hexanoate. The reaction buffer for the immunosensor system was optimized as 0.1 M sodium phosphate (pH 7.4). Concentration-dependent sensor responses were obtained in the vitellogenin concentrations ranging from 0.4864 to 486.4000 nM, with a linear correlation between vitellogenin concentration and frequency shift in double-logarithmic scale. The limit of detection of the immunosensor for carp vitellogenin was presumed as 0.4864 nM.

Photopolymerization and Properties of PCL-Based Biodegradable Molecularly Imprinted Polymers (PCL 기반 생분해성 분자 날인 고분자의 광중합 및 물성)

  • Kim, Sun-Hui;Lee, Kyung-Soo;Kim, Yong-Hoon;Choi, Woo-Jin;Kim, Beom-Soo;Kim, Eung-Kook;Kim, Dae-Su
    • Polymer(Korea)
    • /
    • v.31 no.2
    • /
    • pp.153-159
    • /
    • 2007
  • Biodegradable molecularly imprinted polymers (MIPs) can be applied in the biomedical area of biosensors, drug delivery, etc. Therefore, in this study, biodegradable theophylline MIPs were synthesized via photopolymerization using a poly $(\varepsilon-caprolactone)$ (PCL) macromer as a cross-linker and their physical properties were investigated. The yield for the synthesis of the PCL macromer with terminal acrylate groups was ca. 78 mol%. The products were characterized by the combination of FT-IR and $^1H-NMR$ spectroscopic analyses. UV/Visible spectroscopic analysis for removing and rebinding theophylline was performed by monitoring the theophylline concentration in the solution. In vitro biodegradation tests of the theophylline MIPs performed in phosphate buffered saline (PBS) solution at $37^{\circ}C$ showed good biodegradability of the MIPs.

Selective tyrosine conjugation with a newly synthesized PCB -TE2A-luminol bifunctional chelator

  • Subramani Rajkumar;Hyun Park;Abhinav Bhise;Seong Hwan Cho;Jung Young Kim;Kyo Chul Lee;Jeongsoo Yoo
    • Journal of Radiopharmaceuticals and Molecular Probes
    • /
    • v.7 no.2
    • /
    • pp.85-91
    • /
    • 2021
  • Selective amino acid conjugation of bulky antibodies is a valuable asset for real-time diagnosis and therapy. However, selective conjugation incorporating a chelate-bearing radioactive atom into an antibody without affecting its immunoreactivity is a challenging task. A bifunctional chelator (BFC), a selective amino acid-targeting probe, and a linker have been developed to overcome this problem. Here, we report the synthesis of a novel propylene cross-bridged chelator (PCB)-1,8-N,N'-bis-(carboxymethyl)-1,4,8,11-tetraazacyclotetradecane (TE2A)-luminol BFC via a click reaction and radiolabel it with a 64Cu ion for tyrosine-selective conjugation of trastuzumab. In the initial optimization study, we tried different oxidative addition conditions such as electro-oxidation, hemin, horseradish peroxidase, iodogen tube, chloramine-T, and iodo beads. In this study, up to 82% of 64Cu-PCB-TE2A-luminol was conjugated with the antibody in an iodo bead-catalyzed oxidative addition reaction with an isolated yield of 24.4%.

Polymerization of HEMA by Electron beam Irradiation and Fabrication of Soft contact lens (전자빔조사에 의한 HEMA의 중합과 소프트콘택트렌즈 제조)

  • Hwang, Kwang-Ha;Shin, Joong-Hyeok;Sung, Yu-Jin;Jeong, Keun-Seung;Jun, Jin
    • Journal of Korean Ophthalmic Optics Society
    • /
    • v.17 no.2
    • /
    • pp.135-141
    • /
    • 2012
  • Purpose: Polymerization of HEMA(2-hydroxyethyl methacrylate) which can be used in the soft contact lens has been performed by using electron beam(EB) irradiation, and examined the best condition for the polymerization. Comparing the physical properties of the contact lenses to the one fabricated by thermal polymerization method, we check the use possibility of the EB irradiation to the fabrication of the soft contact lens. Methods: We investigated the degree of polymerization of the HEMA according to the composition of the monomer, the additive ratio and the dose of electron beam (0~120 kGy). The degree of polymerization was measured depending on the EB dose to research the best synthetic condition under the EB irradiation. The physical properties of the contact lens such as water content(%), oxygen transmissibility(Dk/t) and optical transmittance were analysed by using the FT-IR results with comparing the two different polymerization method (thermal and electron beam polymerization) with same additive ratio. Results: When the dose of electron beam was above 100 kGy, the degree of polymerization of HEMA was above 99% with regardless using cross-linker and initiator. The water content of the lens fabricated by EB method showed 10% higher than the one by the thermal method which was 40%. The lens fabricated by EB method also showed higher oxygen transmissibility(Dk/t) as same with the water content, and showed twice higher value in the lens fabricated by pure HEMA. According to the FT-IR results, hydrophilic property of the lens fabricated by EB method was increased due to increasing the intermolecular hydrogen bonding. It showed above 90% optical transmittance in the visible range of wavelength on the contact lenses fabricated by the both of two different polymerization method. Conclusions: The polymerization of HEMA without cross-linker and initiator was successful above 100 kGy of EB irradiation. Moreover the lens fabricated from the polymer synthesized by pure HEMA with 100 kGy of EB showed the highest water content and oxygen transmissibility. Therefore EB irradiation is another possible method to synthesize the polymer which can be used for the soft contact lens.

Development of RGD peptides grafted onto chitosan surfaces; Osteoblast interactions (RGD 펩타이드로 표면개질된 키토산막의 생물학적 영향)

  • Lee, Chang-Kyun;Hwang, Jeong-Hyo;Lee, Yong-Moo;Ku, Young;Rhyu, In-Chul;Lee, Seung-Jin;Han, Soo-Boo;Choi, Sang-Mook;Chung, Chong-Pyoung
    • Journal of Periodontal and Implant Science
    • /
    • v.33 no.1
    • /
    • pp.27-35
    • /
    • 2003
  • 1. 목적 생체재료의 생체친화성을 증진시키고 치유를 촉진하기 위한 목적으로 생체재료의 생화학적 표면개질에 관한 연구가 널리 진행되고 있다. 이와 같은 목적으로 이용되어 온 부착분자에는 아미노산, 펩타이드, 단백질, 효소 및 성장인자들을 들 수 있으며, 이들 분자들을 금속, 골대체물질 및 폴리머와 같은 생체재료의 표면개질에 이용하여 왔다. 이 연구의 목적은 생체적합성이 우수하고 생분해성을 지닌 키토산으로 얇은 막을 제작한 후, 세포외 기질의 구성성분 중 세포부착에 관여하는 RGD 펩타이드를 부착시킨, 표면개질 키토산막의 생물학적 영향을 MG-63 조골양세포를 이용하여 관찰하는 것이다. 2. 방법 2% acetic acid에 키토산 가루를 녹여 만든 2% 키토산 용액으로 24-well 배양접시의 표면을 도포 후 24시간 동안 건조시켜 키토산막을 제작하였다. GRGDS 펩타이드를 cross-linker(EDC, NHS) (Sigma, MO, USA) 용액과 반응시켜서 펩타이드의 카르복실기를 활성화시켰다. 이들을 PBS 완충용액으로 수화시킨 키토산막과 결합시켜 펩타이드의 활성화된 카르복실기와 키토산의 아민기 간에 안정적인 아미드 결합(amide bond)이 형성되도록 하였다. 하루 동안 반응을 일으킨 후 PBS 완충용액과 증류수로 씻어내고 냉동 건조시킴으로써 GRGDS가 결합된 키토산막을 제작하였다. 재료 표면의 화학 성분을 알아보는데 사용되는 방법의 일종인 X-ray photoelectron spectroscopy(XPS) 분석을 통하여 부착분자가 키토산막에 결합된 여부를 확인하였다. GRGDS 펩타이드에 요오드를 결합시킨 후, 이것을 키토산막에 공유 결합시키고 XPS를 통해 요오드가 재료 표면에서 검출되는지를 검사하였다. 요오드가 검출된다면 이것은 키토산막 표면에 실제로 GRGDS 펩타이드가 존재하는 것을 의미하게 된다. 표면개질된 키토산막에 사람조골양세포인 MG-63을 접종하여 이를 실험군으로 하였고, 표면이 개질 되지 않은 키토산막을 대조군으로 하였다. 세포부착의 최적화 농도를 확인하기 위하여 GRGDS를 0.01, 0.05, 0.1, 0.25, 0.5, 1.0mg/ml의 농도로 준비하였다. 배양 후 1일, 7일째에 각 well에서 trypsin EDTA를 이용하여 세포를 분리한 후, 이를 원심 분리하여 세포수측정기를 이용하여 부착 세포의 수를 측정하여 세포의 부착 정도를 비교하였다. 배양 2시간, 24시간 후 주사전자현미경을 이용하여 키토산막에 부착된 세포의 양상을 관찰하였다. 3. 결과 XPS를 통한 표면의 화학 성분 분석 결과 GRGDS 펩타이드를 결합시킨 키토산막에서 요오드가 검출되었으며 펩타이드를 부착하지 않은 대조군에서는 검출되지 않았다. 따라서 cross-linker를 이용한 펩타이드와 키토산막의 공유결합을 확인할 수 있었다. 세포 배양 후 1일째 부착된 세포 수를 측정한 결과 0.1mg/ml 이상의 GRGDS 펩타이드 농도로 공유 결합시킨 키토산막에서 부착 세포 수가 다른 농도에 비해 유의성 있게 많이 관찰되었다. 이 농도 이하에서는 대조군과 실험군간에 세포부착의 유의한 차이가 없었다. 따라서 주사전자현미경을 이용한 부착 세포의 양상에 관한 관찰은 0.1mg/ml 농도의 펩타이드를 이용하였다. 세포 배양 7일째, 부착된 세포 수 측정 결과 GRGDS의 농도에 따른 유의성 있는 차이가 없었으며, 실험군과 대조군간에도 유의성 있는 차이가 없었다. 주사전자현미경 관찰결과 2시간 및 24시간 배양된 실험군 모두에서 별모양의 세포들이 키토산막 표면에 편평하게 잘 부착되어 있으며 많은 위족이 발달된 소견을 보인 반면, 대조군에서는 원형 또는 다각형 모양의 세포들이 실험군에 비해 부착이 덜 되어있는 양상을 보였다. 이 연구를 통하여 기능성 펩타이드를 생체재료의 표면에 공유결합 시키는 방법을 확립할 수 있었으며, RGD 펩타이드의 공유결합으로 표면개질된 키토산막이 조골세포의 부착능을 증진시킬 수 있음을 확인하였다. 표면개질된 생체재료를 소, 중동물에 적용시켜 생체 내에서의 생물학적 영향을 평가할 필요가 있으며, 이 실험의 결과는 향후 다양한 기능성 부착분자를 선발, 고안하여 임플란트용 생체재료의 표면개질에 이용하는 이른바 모방생체재료분야에 널리 활용될 수 있을 것으로 생각된다.

Media Characteristics of PVA-derivative Hydrogels Using a CGA Technique (CGA 제조기법을 응용한 PVA 하이드로젤의 담체 특성)

  • Yoon, Mi-Hae; Kwon, Sung-Hyun;Cho, Dae-Chul
    • Journal of Environmental Science International
    • /
    • v.18 no.3
    • /
    • pp.299-308
    • /
    • 2009
  • We manufactured PVA-derived hydrogels using a foam generation technique that has been widely used to prepare colloidal gas aphrons(CGA). These gels were differentiated to the conventional gels such as for medical or pharmaceutical applications, which have tiny pores and some crystalline structure. Rather these should be used in de-pollution devices or adhesion of cells or biomolecules. The crosslinkers used in this work were amino acid, organic acid, sugars and lipids(vitamins). The structures of the gels were observed in a scanned electron microscope. Amino acids gels showed remarkably higher swelling ratios probably because their typical functional groups help constructing a highly crosslinked network along with hydrogen bonds. Boric acid and starch would catalyze dehydration while structuring to result in much lower water content and accordingly high gel content, leading to less elastic, hard gels. Bulky materials such as ascorbic acid or starch produced, in general, large pores in the matrices and also nicotinamide, having large hydrophobic patches was likely to enlarge pore size of its gels as well since the hydrophobicity would expel water molecules, thus leading to reduced swelling. Hydrophilicity(or hydrophobicity), functional groups which are involved in the reaction or physical linkage, and bulkiness of crosslinkers were found to be more critical to gel's cross linking structure and its density than molecular weights that seemed to be closely related to pore sizes. Microscopic observation revealed that pores were more or less homogeneous and their average sizes were $20{\mu}m$ for methionine, $10-15{\mu}m$ for citric acid, $50-70{\mu}m$ for L-ascorbic acid, $30-40{\mu}m$ for nicotinamide, and $70-80{\mu}m$ for starch. Also a sensory test showed that amino acid and glucose gels were more elastic meanwhile acid and nicotinamide gels turned out to be brittle or non-elastic at their high concentrations. The elasticity of a gel was reasonably correlated with its water content or swelling ratio. In addition, the PVA gel including 20% ascorbic acid showed fair ability of cell adherence as 0.257mg/g-hydrogel and completely degraded phenanthrene(10 mM) in 240 h.

Hydrolysis of Urea by Immobilized Urease Membrane (우레아제(Urease) 고정막에 의한 요소(Urea)의 가수분해)

  • Kim, Byoung-Sik;Kim, Min;Heo, Kwang-Beom;Hong, Joo-Hee;Na, Won-Jae;Kim, Jae-Hun
    • Applied Chemistry for Engineering
    • /
    • v.18 no.1
    • /
    • pp.10-16
    • /
    • 2007
  • In this study, we examined the preparation and hydrolysis property of immobilized urease membrane to decompose harmful urea in the body and remove ammonia which was produced by its decomposition. Urease immobilized membrane was prepared by introducing anion-exchange group DEA into porous hollow-fiber membrane by radiation graft polymerization method, and immobilization of urease. When urease was immobilized at membrane introduced with anion-exchange group, the more increasing grafting rate, the more increasing immobilization amount. The result originates from the fact that a greater amount of protein was immobilized by forming a multilayer on the longer grafted chain. Meanwhile, the addition of the cross-linker was possible not only to suppress separation phenomenon produced during a washing process of immobilized urease membrane but also to enable the recycling of membrane. Urease Immobilized membrane with no separation phenomenon was prepared by cross-linking reaction for 5 h, and the hydrolysis rate of prepared urease immobilized membrane was over 98% and 50%, respectively, in 1 mol and 4 mol urea solutions.

ELISA Development for the residue of the organophosphorus insecticide acephate (ELISA에 의한 유기인계 살충제 Acephate 잔류물 분석법 개발)

  • Lee, Jae-Koo;Ahn, Ki-Chang;Stoutamire, Donald W.;Gee, Shirley J.;Hammock, Bruce D.
    • The Korean Journal of Pesticide Science
    • /
    • v.5 no.2
    • /
    • pp.1-12
    • /
    • 2001
  • A competitive indirect enzyme-linked immunosorbent assay (ci ELISA) for the organophosphorus insecticide acephate, O,S-dimethyl acetylphosphoramidothioate, was developed using a polyclonal antibody. Three different haptens mimicking the analyze and containing hexanoic acid moiety as a linker were synthesized, and then conjugated with the carrier proteins bovine serum albumin and keyhole limpet hemocyanin by the N-hydroxysuccinimide active ester method. Polyclonal antibodies raised against hapten-KLH conjugates in rabbits and the hapten-BSA conjugates as coating antigens were screened and selected for the assay in the homologous and/or heterologous ELISA system. The effects of various assay conditions, including blocking reagents, detergent content, organic solvents, pH, and preincubation of tile mixture of the polyclonal antibody and the analyze on the sensitivity were evaluated. The $IC_{50}$ value of acephate of 110 ng/mL was obtained in an optimized heterologous system using hapten-3-BSA as a coating antigen and a polyclonal antibody 8377, showing the detection range of 10-1000 ng/mL and the lowest detection limit of 4 ng/mL. The cross-reactivities of the structurally related insecticides, including methamidophos were less than 0.02%. These results indicate that the ELISA could be a convenient and alternative tool for monitoring acephate residues in agricultural products and environmental samples.

  • PDF

Current Research Trends on Surface Modification of Pressure-driven Membranes for Fouling Mitigation (압력 구동 기반 분리막의 막 오염 저감을 위한 표면 개질 방법 최신 연구 동향)

  • Jun, Byung-Moon;Lee, Hyung Kae;Kim, Woo Jeong;Park, Jihun;Kim, Jong Hyeok;Kwon, Young-Nam
    • Membrane Journal
    • /
    • v.28 no.1
    • /
    • pp.1-20
    • /
    • 2018
  • Fresh water is an important resource for humans, and pressure-driven membrane technology has been widely known as an energy-efficient method to obtain water resource. However, membrane fouling phenomenon, which is one of the major issue during operation, deteriorates membrane permeability. These fouling is usually affected by interaction between surface of membrane and various foulants, therefore, modification of membrane's surface is one of the methods to improve fouling-resistance. This review focuses on the method to modify surface of pressure-driven membranes such as microfiltration (MF), ultrafiltration (UF), nanofiltration (NF), and reverse osmosis (RO). Specifically, there are two different surface modification methods: (1) adsorption and coating as the physical modification methods, (2) cross-linker, free radical polymerization (FRP), atom transfer radical polymerization (ATRP), plasma/UV-induced polymerization as the chemical modification methods. This review introduces the physico - chemical surface modification methods reported in recent papers and suggests research directions for membrane separation which can increase membrane fouling resistance.

Characterization of Hydrogel Tinted Contact Lens Containing 4-iodoaniline using Titanium Silicon Oxide Nanoparticles as Additive (티타늄 실리콘 옥사이드 나노입자를 첨가제로 사용한 4-iodoaniline을 포함한 하이드로젤 착색 콘택트렌즈의 특성)

  • Cho, Seon-Ahr;Sung, A-Young
    • Journal of Korean Ophthalmic Optics Society
    • /
    • v.19 no.3
    • /
    • pp.315-322
    • /
    • 2014
  • Purpose: The physical and optical characteristics of hydrophilic tinted contact lens containing titanium silicon oxide nanoparticles and the basic hydrogel contact lens material containing 4-iodoaniline were examined. In this study, the utility of titanium silicon oxide nanoparticles as a UV-blocking material for ophthalmologic devices were investigated by measuring the UV transmittance of the produced polymer. Also, titanium silicon oxide nanoparticles only without the addition of 4-iodoaniline in primary contact lens materials by copolymerizing two groups were compared. Methods: For manufacturing hydrogel lens, HEMA, MA, MMA, 4-iodoaniline and a cross-linker EGDMA were copolymerized in the presence of AIBN as an initiator. Also, the titanium silicon oxide nanoparticles was used as additive. After polymerization the physical properties such as water content, refractive index, contact angle and spectral transmittance of produced contact lenses were measured. Results: Measurement of the physical properties of the copolymerized material showed that the water content, refractive index, UV-B transmittance and contact angle were in the range of 35.01~38.60%, 1.4350~1.4418, $34.15{\sim}57.25^{\circ}$ and 1.0~10.0%, respectively. Titanium silicon oxide nanoparticles is not used as an additive in the experimental group, the results of the measurement showed that the water content, refractive index, contan angle and UV-B transmittance of the hydrogel lens polymer was 34.00~36.80%, 1.4378~1.4420, $40.15{\sim}60.16^{\circ}$ and 1.8~25.0%, respectively. Conclusions: Also, the transmittance for UV light was reduced significantly in combinations containing titanium oxide nanoparticles.