• Title/Summary/Keyword: Automated liquid-handling system

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

  • Shin, Sun-Young;Kim, Hee-Sun;Lee, Hyun-Ju;Jang, Hyun-Young;Woo, Jae-Ryong;Lee, Ho-Young
    • The Korean Journal of Nuclear Medicine Technology
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    • v.14 no.2
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    • pp.190-196
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    • 2010
  • Purpose: This study was undertaken to investigate the feasibility of using automated liquid-handling systems equipped with reusable fixed tips in Radioimmunoassays and to demonstrate that the use of an automated pipetting instrument can decrease in the typical daily workload. Materials and Methods: The precisions of the automated pipettor and the manual pipettor were determined gravimetrically (n=30). A total of 30 specimens for HBs Ag were repeatedly pipetted (three replicates) with the automated pipettor and then retested. PSA samples were simultaneously pipetted with either the automated pipettor or the manual pipettor and then analyzed (n=40). Sample carryover test assessed for CA19-9, AFP and HCG when the automated pipettor was used. Pipetting speed of the automated pipettor and the manual pipettor were compared by evaluation of each workload. Results: The precisions coefficients of variation (CV) were 2.1% for the automated pipettor and 1.6% for manual pipettor. The mean cpm and CV for each group of replicates were 41,203 cpm and 3.7% for HBs Ag positive specimens, and 99 cpm and 7.9% for HBs Ag negative specimens, respectively. PSA results showed no significant differences between automated pipettor and manual pipettor (p=0.15, r=0.999). Carryover for CA19-9, AFP and HCG analytes was <0.1 ppm or below the assay limit of detection. Pipetting speed was significantly improved by using the automated instrument. Conclusion: There was no evidence that the use of an automated pipettor adversely affected any of the performance characteristics of the assay. Indeed, routine use of the Tecan automated pipettor has resulted in a decrease in the typical daily workload.

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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
    • Proceedings of the PSK Conference
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    • 2003.10b
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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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    • v.15 no.1
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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.

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

  • Chung, Jae-Woo;Kim, Pan-Soo
    • Journal of Korean Society of Industrial and Systems Engineering
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    • v.34 no.3
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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
    • Proceedings of the Korean Society of Toxicology Conference
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    • 2006.11a
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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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    • 2003.10a
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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
    • Biomedical Science Letters
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    • v.29 no.4
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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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    • v.26 no.5
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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.