• 제목/요약/키워드: Automated liquid-handling system

검색결과 8건 처리시간 0.025초

고정용 팁을 사용하는 Automated Liquid-Handling System의 RIA적용의 평가 (Evaluation of an Automated Liquid-Handling System with Fixed Pipetting Tips in Radioimmunoassay)

  • 신선영;김희선;이현주;장현영;우재룡;이호영
    • 핵의학기술
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    • 제14권2호
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    • pp.190-196
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    • 2010
  • RIA검사에 재생 가능한 고정용 팁이 장착된 Automated Luquid-Handling System을 적용하여 그 사용 가능성을 조사하고, 기존의 single-pipetting과 비교하여multi-pipetting을 이용한 자동분주기의 사용이 검사자의workload를 감소시켜 업무효율을 증가시키는 것을 확인하고자 하는 것이다. Multipipetting을 이용한 자동화장비를 RIA에 적용함으로써 검사 결과의 정확도와 정밀도를 높이고, 검사횟수를 증가시켜 당일진료체계를 구축하여 오늘날의 의료 현장의 요구를 충족 시키고자 한다. 고정된 팁이 장착된 Automated pipettor의 RIA적용 가능성을 조사하기 위하여 분주량의 정확도와 정밀도, 검사결과의 재현성, 일치도, 그리고 검체간 이월오염을 측정하였다. Multi-pipetting을 이용한 자동분주기의 사용이 workload를 감소시키는 것을 시연하기 위하여 daily workload를 20일 동안 측정하였다. Tecan automated pipettor의 정밀도를 측정하였는데, Tecan은 평균 2.1%, 그리고 manual pipettor는 평균 1.6%의 CV를 보였다. Tecan automated pipettor의 재현성을 평가하여 반복 측정된 각각의 그룹에 대한 cpm결과의 평균과 CV를 계산하였는데, HBs Ag 양성 검체는 41,203 cpm과 3.7%의 CV를 보였다. HBs Ag 음성 검체는 99 cpm과 7.9%의 CV를 보였다. 일치도 평가에서는 Tecan automated pipettor 와 Gilson manual pipettor로 분주하여 결과를 분석하였는데, 측정된 결과는 p값이 0.150로 유의한 차가 나타나지 않았고, r값은 0.999로 높은 상관관계를 보였다. 검체간 이월오염은 1 ppm이하임을 확인하였다. 본 연구는 Tecan Freedom evo 100 같은 automated pipetting 장비를 RIA검사에 사용하였을 때, 검사결과의 질을 떨어뜨리지 않으면서 검사의 효율을 향상시킨다는 것을 시연하였다. 자동화장비의 사용은 증가하는 검사건수에 대한 대안이 될 것이며, 검사건수가 증가하더라도 신속하고 정확한 검사결과를 보고 할 수 있게 될 것이다.

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High throughput automated 96-well solid-phase extraction and liquid chromatography-tandem mass spectrometric analysis of beraprost in human plasma

  • Chang, Kyu-Young;Kim, Ho-Hyun;Lee, Hee-Joo;Lee, Kyung-Ryul
    • 대한약학회:학술대회논문집
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    • 대한약학회 2003년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.2-2
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    • pp.224.1-224.1
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    • 2003
  • A sensitive and selective liquid chromatographic method coupled with tandem mass spectrometry (LC-MS/MS) was developed for the determination of beraprost in human plasma. Plasma samples were transferred into 96-well OASIS HLB extraction plate using an automated sample handling system and the drugs were eluted with methanol. The eluents were then evaporated and reconstituted with water. All sample transfer and solid-phase extraction (SPE) was automated through the application of both the PerkinElmer MultiPROBE II HT and TOMTEC Quadra 96 workstation. (omitted)

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Optimization of Automated Suspension Trapping Digestion in Bottom-Up Proteomics via Mass Spectrometry

  • Haneul Song;Yejin Jeon;Iyun Choi;Minjoong Joo;Jong-Moon Park;Hookeun Lee
    • Mass Spectrometry Letters
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    • 제15권1호
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    • pp.62-68
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    • 2024
  • The Suspension Trapping (S-Trap) method has been a prominent sample preparation technique since its introduction in 2014. Its capacity to induce protein aggregation using organic solvents has significantly improved protein purification and facilitated peptide identification. However, its full potential for automation has been limited by the lack of a suitable liquid handling system until recently. In this study, we aimed to enhance the automation of S-Trap sample preparation by optimizing the S-Trap digestion process, incorporating triethylammonium bicarbonate (TEAB) and CaCl2. The utilization of TEAB buffer conditions in this innovative process led to a noteworthy 12% improvement in protein identification. Additionally, through careful observation of various incubation conditions, we streamlined the entire sample preparation workflow into a concise 4 hours timeline, covering reduction, alkylation, and trypsin incubation stages. This refined and expedited automated S-Trap digestion process not only showcased exceptional time efficiency but also improved trypsin digestion, resulting in increased protein identification.

LCD공정에서 스토커시스템 성과측정 모델 (A Performance Model for Stocker Systems in Liquid Crystal Display (LCD) Fabrication Lines)

  • 정재우;김판수
    • 산업경영시스템학회지
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    • 제34권3호
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    • pp.1-7
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    • 2011
  • The stocker system is another name of automated storage and retrieval system (AS/RS) and being popularly used as main material handling tools in Liquid Crystal Display (LCD) and semiconductor fabrication facilities. Recently the use of the stocker system has been extended to transportation from conventional storage and retrieval in LCD fabrication facilities. Toolsets are connected in the ground level of the stocker system and 4~6 stories of the shelves are placed in the upper or lower ground level. As a consequence of the more sophisticated design, move requests imposed on the system greatly increased. For solving this problem, the industry adopted the dual-robot stocker system that two robots are moving along the same guide line in the stocker system. This research develops a closed-form solution to estimate a delivery rate of the dual robot stocker system under given design and operation parameters. Using this stochastic model, industry practitioners could analyze performance levels under given various design parameters, and ultimately the model helps optimizing the design parameters.

Full validation of high-throughput bioanalytical method for the new drug in plasma by LC-MS/MS and its applicability to toxicokinetic analysis

  • Han, Sang-Beom
    • 한국독성학회:학술대회논문집
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    • 한국독성학회 2006년도 추계학술대회
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    • pp.65-74
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    • 2006
  • Modem drug discovery requires rapid pharmacokinetic evaluation of chemically diverse compounds for early candidate selection. This demands the development of analytical methods that offer high-throughput of samples. Naturally, liquid chromatography / tandem mass spectrometry (LC-MS/MS) is choice of the analytical method because of its superior sensitivity and selectivity. As a result of the short analysis time(typically 3-5min) by LC-MS/MS, sample preparation has become the rate- determining step in the whole analytical cycle. Consequently tremendous efforts are being made to speed up and automate this step. In a typical automated 96-well SPE(solid-phase extraction) procedure, plasma samples are transferred to the 96-well SPE plate, internal standard and aqueous buffer solutions are added and then vacuum is applied using the robotic liquid handling system. It takes only 20-90 min to process 96 samples by automated SPE and the analyst is physically occupied for only approximately 10 min. Recently, the ultra-high flow rate liquid chromatography (turbulent-flow chromatography)has sparked a huge interest for rapid and direct quantitation of drugs in plasma. There is no sample preparation except for sample aliquotting, internal standard addition and centrifugation. This type of analysis is achieved by using a small diameter column with a large particle size(30-5O ${\mu}$m) and a high flow rate, typically between 3-5 ml/min. Silica-based monolithic HPLC columns contain a novel chromatographic support in which the traditional particulate packing has been replaced with a single, continuous network (monolith) of pcrous silica. The main advantage of such a network is decreased backpressure due to macropores (2 ${\mu}$m) throughout the network. This allows high flow rates, and hence fast analyses that are unattainable with traditional particulate columns. The reduction of particle diameter in HPLC results in increased column efficiency. use of small particles (<2 urn), however, requires p.essu.es beyond the traditional 6,000 psi of conventional pumping devices. Instrumental development in recent years has resulted in pumping devices capable of handling the requirements of columns packed with small particles. The staggered parallel HPLC system consists of four fully independent binary HPLC pumps, a modified auto sampler, and a series of switching and selector valves all controlled by a single computer program. The system improves sample throughput without sacrificing chromatographic separation or data quality. Sample throughput can be increased nearly four-fold without requiring significant changes in current analytical procedures. The process of Bioanalytical Method Validation is required by the FDA to assess and verify the performance of a chronlatographic method prior to its application in sample analysis. The validation should address the selectivity, linearity, accuracy, precision and stability of the method. This presentation will provide all overview of the work required to accomplish a full validation and show how a chromatographic method is suitable for toxirokinetic sample analysis. A liquid chromatography/tandem mass spectrometry (LC-MS/MS) method developed to quantitate drug levels in dog plasma will be used as an example of tile process.

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Automated Protein-Expression Profiling System using Crude Protein Direct Blotting Method

  • Kobayashi, Hironori;Torikoshi, Yasuhiro;Kawasaki, Yuko;Ishihara, Hideki;Mizumoto, Hiroshi
    • 제어로봇시스템학회:학술대회논문집
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    • 제어로봇시스템학회 2003년도 ICCAS
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    • pp.2356-2361
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    • 2003
  • Proteome research in the medical field is expected to accelerate the understanding of disease mechanism, and to create new diagnostic concept. For protein profiling, this paper proposes a new methodology named CPDIB (Crude Protein Direct Blotting). In the CPDIB procedure, crude protein sample is directly immobilized on a membrane and the expression of protein molecules in the sample are analyzed quantitatively by using a special device called ImmobiChip, where the membrane is used as a field of the immune reaction. The over-all structure of the ImmobiChip is based on the conventional Slot blot device. Mechanical improvement in the air-tightness of the case holding the membrane realizes the direct blotting and results in high performance of stability in the immune reaction. In the measurement of multiple proteins, a dispensing robot is used for increasing the efficiency of handling of liquid. Cooperation of the dispensing robot with the ImmobiChip for immobilizing proteins realizes automated and stable performance of the CPDIB procedure. This paper shows the evaluation of the air-tightness of the ImmobiChip, the ability of analyzing proteins using the CPDIB procedure and the performance of the automated equipment.

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Development of an Automatic PCR System Combined with Magnetic Bead-based Viral RNA Concentration and Extraction

  • MinJi Choi;Won Chang Cho;Seung Wook Chung;Daehong Kim;Il-Hoon Cho
    • 대한의생명과학회지
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    • 제29권4호
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    • pp.363-370
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    • 2023
  • Human respiratory viral infections such as COVID-19 are highly contagious, so continuous management of airborne viruses is essential. In particular, indoor air monitoring is necessary because the risk of infection increases in poorly ventilated indoors. However, the current method of detecting airborne viruses requires a lot of time from sample collection to confirmation of results. In this study, we proposed a system that can monitor airborne viruses in real time to solve the deficiency of the present method. Air samples were collected in liquid form through a bio sampler, in which case the virus is present in low concentrations. To detect viruses from low-concentration samples, viral RNA was concentrated and extracted using silica-magnetic beads. RNA binds to silica under certain conditions, and by repeating this binding reaction, bulk samples collected from the air can be concentrated. After concentration and extraction, viral RNA is specifically detected through real-time qPCR (quantitative polymerase chain reaction). In addition, based on liquid handling technology, we have developed an automatic machine that automatically performs the entire testing process and can be easily used even by non-experts. To evaluate the system, we performed air sample collection and automated testing using bacteriophage MS2 as a model virus. As a result, the air-collected samples concentrated by 45 times then initial volume, and the detection sensitivity of PCR also confirmed a corresponding improvement.

Simultaneous Characterization of Sofalcone and Its Metabolite in Human Plasma by Liquid Chromatography -Tandem Mass Spectrometry

  • Han, Sang-Beom;Jang, Moon-Sun;Lee, Hee-Joo;Lee, Ye-Rie;Yu, Chong-Woo;Lee, Kyung-Ryul;Kim, Ho-Hyun
    • Bulletin of the Korean Chemical Society
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    • 제26권5호
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    • pp.729-734
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    • 2005
  • A sensitive and selective method for quantitation of sofalcone and its active metabolite in human plasma has been established using liquid chromatography-electrospray ionization tandem mass spectrometry (LC-ESI/MS/MS). Plasma samples were transferred into 96-well plate using an automated sample handling system and spiked with 10 $\mu$L of 2 $\mu$g/mL $d_3$-sofalcone and $d_3$-sofalcone metabolite solutions (internal standard), respectively. After adding 0.5 mL of acetonitrile to the 96-well plate, the plasma samples were then vortexed for 30 sec. After centrifugation, the supernatant was transferred into another 96-well plate and completely evaporated at 40 ${^{\circ}C}$ under a stream of nitrogen. Dry residues were reconstituted with mobile phase and were injected into a $C_{18}$ reversed-phase column. The limit of quantitation of sofalcone and its metabolite was 2 ng/mL, using a sample volume of 0.2 mL for analysis. The reproducibility of the method was evaluated by analyzing 10 replicates over the concentration range of 2 ng/mL to 1000 ng/mL. The validation experiments of the method have shown that the assay has good precision and accuracy. Sofalcone and its metabolite produced a protonated precursor ion ([M+H]$^+$) of m/z 451 and 453, and a corresponding product ion of m/z 315 and 317, respectively. Internal standard ($d_3$-sofalcone and $d_3$-sofalcone metabolite) produced a protonated precursor ion ([M+H]$^+$) of m/z 454 and 456 and a corresponding product ion of m/z 315 and 317, respectively. The method has been successfully applied to a pharmacokinetic study of sofalcone and its active metabolite in human plasma.