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Construction of a Large Synthetic Human Fab Antibody Library on Yeast Cell Surface by Optimized Yeast Mating

  • Baek, Du-San;Kim, Yong-Sung
    • Journal of Microbiology and Biotechnology
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    • v.24 no.3
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    • pp.408-420
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    • 2014
  • Yeast surface-displayed antibody libraries provide an efficient and quantitative screening resource for given antigens, but suffer from typically modest library sizes owing to low yeast transformation efficiency. Yeast mating is an attractive method for overcoming the limit of yeast transformation to construct a large, combinatorial antibody library, but the optimal conditions have not been reported. Here, we report a large synthetic human Fab (antigen binding fragment) yeast surface-displayed library generated by stepwise optimization of yeast mating conditions. We first constructed HC (heavy chain) and LC (light chain) libraries, where all of the six CDRs (complementarity-determining regions) of the variable domains were diversified mimicking the human germline antibody repertoires by degenerate codons, onto single frameworks of VH3-23 and $V{\kappa}1$-16 germline sequences, in two haploid cells of opposite mating types. Yeast mating conditions were optimized in the order of cell density, media pH, and cell growth phase, yielding a mating efficiency of ~58% between the two haploid cells carrying HC and LC libraries. We constructed two combinatorial Fab libraries with CDR-H3 of 9 or 11 residues in length with colony diversities of more than $10^9$ by one round of yeast mating between the two haploid HC and LC libraries, with modest diversity sizes of ${\sim}10^7$. The synthetic human Fab yeast-displayed libraries exhibited relative amino acid compositions in each position of the six CDRs that were very similar to those of the designed repertoires, suggesting that they are a promising source for human Fab antibody screening.

Manufacturing and Characterization of PVDF/TiO2 Composite Nano Web with Improved β-phase (β-phase가 향상된 PVDF/TiO2 Nano Web 제조 및 특성 분석)

  • Bae, Sung Jun;Kim, Il Jin;Lee, Jae Yeon;Sur, Suk-Hun;Choi, Pil Jun;Sim, Jae Hak;Lee, Seung Geol;Ko, Jae Wang
    • Textile Coloration and Finishing
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    • v.32 no.3
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    • pp.167-175
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    • 2020
  • In this study, the optimum conditions for manufacturing PVDF nano web according to various electrospinning conditions such as solution concentration and applied voltage conditions were confirmed. The optimum spinning conditions were studied by analyzing the changes in the radioactivity of PVDF/TiO2 nano web according to the TiO2 content and the content of β-phase closely related to the piezoelectric properties under established conditions. As a result, it was confirmed that the concentration of the spinning solution was 20 wt%, the applied voltage was 25 kV, and the TiO2 content was 5 phr. PVDF nano web and PVDF/TiO2 nano web were observed morphologies through Scanning Electron Microscope(SEM) analysis. Formation of β-phase by electrospinning was confirmed by Fourier transform infrared spectroscopy(FT-IR) and X-ray Diffractometer(XRD), and the effect of the trapped nano web structure on the piezoelectric properties was investigated.

Effect of Propofol, an Intravenous Anesthetic Agent, on $K_{ATP}$ Channels of Pancreatic ${\beta}-cells$ in Rats

  • Park, Eun-Jee;Song, Dae-Kyu;Cheun, Jae-Kyu;Bae, Jung-In;Ho, Won-Kyung;Earm, Yung-E
    • The Korean Journal of Physiology and Pharmacology
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    • v.4 no.1
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    • pp.25-31
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    • 2000
  • ATP-sensitive potassium channels ($K_{ATP}$ channels) play an important role in insulin secretion from pancreatic beta cells. We have investigated the effect of propofol on $K_{ATP}$ channels in cultured single pancreatic beta cells of rats. Channel activity was recorded from membrane patches using the patch-clamp technique. In the inside-out configuration bath-applied propofol inhibited the $K_{ATP}$ channel activities in a dose-dependent manner. The half-maximal inhibition dose (ED50) was $48.6{\pm}8.4\;{\mu}M$ and the Hill coefficient was $0.73{\pm}0.11.$ Single channel conductance calculated from the slope of the relationship between single channel current and pipette potential $(+20{\sim}+100\;mV)$ was not significantly altered by propofol $(control:\;60.0{\pm}2.7\;pS,\;0.1\;mM\;propofol:\;58.7{\pm}3.5\;pS).$ However, mean closed time was surely increased. Above results indicate that propofol blocks the $K_{ATP}$ channels in the pancreatic beta cells in the range of its blood concentrations during anesthesia, suggesting a possible effect on insulin secretion and blood glucose level.

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Examination on the Mounting Status of Cigar Lighter Receptacle for Vehicles and Analysis of its Tracking Characteristics (차량용 시가 잭의 장착 실태조사 및 트레킹 특성 분석)

  • Choi, Chung-Seog
    • Journal of the Korean Society of Safety
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    • v.24 no.4
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    • pp.28-33
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    • 2009
  • This study examined the mounting status of cigar lighter receptacles for vehicles and analyzed the tracking phenomenon that occurs when foreign material entered a cigar lighter receptacle to obtain data for the analysis of accident investigation. Regardless of the vehicle's output, cigar lighter receptacles are mounted in a vehicle horizontally, vertically, or at tilting or inclined angle. The tilting type cigar lighter receptacle is much easier to use but current leakage resulting from foreign materials (coffee, beverages, water, etc.) falling into the cigar lighter receptacle may cause a fire to start. This study used a vehicle battery (DC 12V) as a power supply for the tracking test and configured its circuit in the same way as that of an electrical device in a vehicle. The tracking phenomenon that occurred in the standby mode of the vehicle exhibited a fine flame and an irregular occurrence of smoke. While this tracking phenomenon was occurring, the leakage current and the reaching distance of the flame were measured to be approximately 930mA and $20{\sim}50cm$, respectively. It is thought that the resultant flame may ignite toluene, dust, cigarettes, etc. It was observed that as the tracking progressed, the internal metal socket melted and a hole was created, the surface of which was also severely carbonized. In addition, the electrical resistance of the carbonized conductive path was measured to be approximately $30{\Omega}$. It is thought that this much resistance may cause local heating when leakage current flows and could ignite any nearby flammable material.

입자침전법을 이용한 광도전체 필름의 X선 반응 특성에 관한 연구

  • Choe, Chi-Won;Gang, Sang-Sik;Jo, Seong-Ho;Gwon, Cheol;Nam, Sang-Hui
    • Proceedings of the Korean Institute of Electrical and Electronic Material Engineers Conference
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    • 2007.06a
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    • pp.176-176
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    • 2007
  • Flat-panel direct conversion detectors used in compound substance of semiconductor are being studied for digital x-ray imaging. Recently, such detectors are deposited by physical vapor deposition(PVD) generally. But, most of materials (HgI2, PbI2, TlBr, PbO) deposited by PVD have shown difficult fabrication and instability for large area x-ray imaging. Consequently, in this paper, we propose applicable potentialities for screen printing method that is coated on a substrate easily. It is compared to electrical properties among semiconductors such as $HgI_2$, $PbI_2$, PbO, HgBrI, InI, and $TlPbI_3$ under investigation for direct conversion detectors. Each film detector consists of an ~25 to $35\;{\mu}m$ thick layer of semiconductor and was coated onto the substrate. Substrates of $2cm{\times}2cm$ have been used to evaluate performance of semiconductor radiation detectors. Dark current, sensitivity and physics properties were measured. Leakage current of $HgI_2$ as low as $9pA/mm^2$ at the operation bias voltage of ${\sim}1V/{\mu}m$ was observed. Such a value is not better than PVD process, but it is easy to be fabricated in high quality for large area x-ray Imaging. Our future efforts will concentrate on optimization of growth of film thickness that is coated onto a-Si TFT array.

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Hydrophilic property by contact angle change of ion implanted polycarbonate (이온주입 Polycarbonate의 접촉각 변화에 의한 친수특성)

  • Lee, Chan-Young;Lee, Jae-Hyung;Lim, Kee-Joe
    • Proceedings of the Korean Institute of Electrical and Electronic Material Engineers Conference
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    • 2003.11a
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    • pp.533-538
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    • 2003
  • It has been shown that ion implantation produces remarkable improvements in surface-sensitive physical and chemical properties as well as other mechanical properties, in polymers. In this study, ion implantation was performed onto polymer, PC(polycarbonate), in order to investigate surface hydrophilic property through contact angle measurement using distilled water. PC was irradiated with N, Ar, Xe ions at the irradiation energy of $20\;{\sim}\;50keV$ and the dose range of $5{\times}10^{15},\;1{\times}10^{16},\;7{\times}10^{16}\;ions/cm^2$. The contact angle of water has been reduced with increasing fluence and ion mass but increased with increasing implanted energy. The changes of chemical and structural property are discussed in view of infrared spectroscopy and FT-IR, XPS, which shows increasing C-O bonding and C-C bonding. The root mean square of surface roughness examined by means of AFM changed smoothly from 0.387nm to 0.207nm and the change of wettability was discussed with respect to elastic and inelastic collisions obtained as results of TRIM simulation. It was found that wettability of the modified PC surface was affected on change of functional group and nuclear stopping or linear energy transfer(LET, energy deposited per unit track length per ion) that causes chain scission by displacing atom from polymer chains, but was not greatly dependant on surface morphology.

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Fabricated thin-film transistors with P3HT channel and $NiO_x$ electrodes (P3HT와 IZO 전극을 이용한 thin film transistors 제작)

  • Kang, Hee-Jin;Han, Jin-Woo;Kim, Jong-Yeon;Moon, Hyun-Chan;Park, Gwang-Bum;Kim, Tae-Ha;Seo, Dae-Shik
    • Proceedings of the Korean Institute of Electrical and Electronic Material Engineers Conference
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    • 2006.06a
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    • pp.467-468
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    • 2006
  • We report on the fabrication of P3HT-based thin-film transistors (TFT) that consist of indium-zinc-oxide (IZO), PVP (poly-vinyl phenol), and Ni for the source-drain (S/D) electrode, gate dielectric, and gate electrode, respectively. The IZO S/D electrodes of which the work function is well matched to that of P3HT were deposited on a P3HT channel by thermal evaporation of IZO and showed a moderately low but still effective transmittance of ~25% in the visible range along with a good sheet resistance of ${\sim}60{\Omega}/{\square}$. The maximum saturation current of our P3HT-based TFT was about $15{\mu}A$ at a gate bias of -40V showing a high field effect mobility of $0.05cm^2/Vs$ in the dark, and the on/off current ratio of our TFT was about $5{\times}10^5$. It is concluded that jointly adopting IZO for the S/D electrode and PVP for gate dielectric realizes a high-quality P3HT-based TFT.

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Temperature Dependence of Volume Resistivity on Epoxy Nano-composites (에폭시 나노컴퍼지트 체적 고유저항의 온도 의존성)

  • Kim, Chang-Hoon;Lee, Young-Sang;Kang, Yong-Gil;Park, Hee-Doo;Shin, Jong-Yeol;Hong, Jin-Woong
    • Journal of the Korean Institute of Electrical and Electronic Material Engineers
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    • v.24 no.10
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    • pp.834-838
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    • 2011
  • This research shows the electrical characteristic using excellent epoxy nano-composite of MgO 5.0 wt% and $SiO_2$ 0.4 wt% in mechanical strength test depending on nano-additive. First of all, volume resistance depending on nano-additive and temperature using high resistance meter (HP. 4329A) by increasing 10, 100, 1,000 V of applying voltage was measured. Moreover, temperature range of $25{\sim}120^{\circ}C$ with virgin sample was tested using TO-9B oven by Ando Company. The result showed that virgin and the samples added with MgO and $SiO_2$ had similar value of volume resistance in low temperature and low electric field region and reduced with slow slope. The nano-composite's volume resistance of sample added with MgO and $SiO_2$ had higher value than virgin sample's volume resistance in high temperature region more than $80^{\circ}C$. Moreover, the slope has steeply reduced. The volume resistance of sample added with MgO 5.0 wt% was $8.38{\times}10^{13}\;{\Omega}{\cdot}cm$ and it was 6.8 times more than virgin sample in high temperature at $120^{\circ}C$. The insulation characteristics were constant although filler has changed in low temperature region. But, in high temperature region, the value of volume resistance of sample with MgO 5.0 wt% was 7.6 times more than the virgin sample's volume resistance.

Change in Electrical Properties of Al2O3/GaN MIS Structures according to the Thickness of Al2O3 Thin Film and Annealing Temperature (산화알루미늄 박막의 두께 및 열처리 온도에 따른 Al2O3/GaN MIS 구조의 전기적 특성 변화)

  • Kwak, No-Won;Lee, Woo-Seok;Kim, Ka-Lam;Kim, Hyun-Jun;Kim, Kwang-Ho
    • Journal of the Korean Institute of Electrical and Electronic Material Engineers
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    • v.22 no.6
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    • pp.470-475
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    • 2009
  • We deposited $Al_2O_3$ thin films on GaN by remote plasma atomic layer deposition (RPALD) technique, trimethylaluminum(TMA) and oxygen were used as precursors, at fixed process condition, the number of cycle were changed. Growth rate per cycle was $1.2\;{\AA}$/cycle. and Growth rate was in proportion to a number of cycle, the GaN MIS capacitors that $Al_2O_3$ thin film were deposited above 12 nm, have excellent electrical properties, a low electrical leakage current density(${\sim}10^{-10}\;A/cm^2$ at 1.5 MV), but below 12 nm, we can see the degradation of the leakage current density. After post deposition annealing, Dielectric constant was estimated by 1 MHz high-frequency C-V method, it was varied with the anealing temperature from 6.9 at no post anealed to 7.6 at $800^{\circ}C$, and we can see a improvement of the leakage current density and breakdown voltage by post deposition anealing below $700^{\circ}C$, but, after anealed at $800^{\circ}C$, we can see the degradation of the leakage current density and breakdown voltage.

A Study on the Additives of mixed Gas charged in Thermostatic Bulb for Expansion Valve (팽창밸브 개폐용 감온통 혼합가스의 첨가제 연구)

  • Kim, Si-Young;Ju, Chang-Sik;Koo, Su-Jin
    • Journal of Power System Engineering
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    • v.18 no.6
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    • pp.126-132
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    • 2014
  • The P-T characteristics of mixed refrigerant in thermostatic expansion valve sensing bulb were studied using R-134a and R-410A refrigerant. The characteristics of mixed refrigerant were investigated according to pressure variation and the variation of composition ratio of R-134A and R-410A in the temperature range of $-15^{\circ}C{\sim}15^{\circ}C$. The Thermodynamic characteristic values of the mixed refrigerants were identified using the characteristic value analysis program of mixed refrigerant(Refrop v9.0, NIST). The P-T characteristics in the case of the mixing ratio of 90:10 for R-410A and R-134A were the same result as R-22. And the physical properties showed similar results with R-22. The Maximum operating pressure(MOP) of mixed refrigerant showed a tendency to decrease with decreasing the mixing ratio of additive gases($N_2$ or He) gases. The characteristics in the case of the mixing ratio of 80:1 for mixed refrigerant and additive gases were the similar result as Reference refrigerant.(R-22 MOP, Sporlan company) In addition $N_2$ and He, both showed the same results. It was able to confirm that a MOP on the thermostatic expansion valve sensing bulb can be maintained by adjusting the mixing ratio of mixed refrigerant gases and additive gases.