• Title/Summary/Keyword: MALDI-TOF-TOF

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The study for the synthesis and analysis of metal chelated protein (단백질의 메탈 킬레이션 화합물 제조 및 확인에 관한 연구)

  • Kim, Sung-Ho;Nam, Hae-Seon;Lee, Yoon-Jin
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.8 no.5
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    • pp.1273-1278
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    • 2007
  • In living system, various types of metal chelated protein show their intrinsic biological activities. In order to investigate the chelation reaction between protein and metal, zinc chelated peptide was systhesized by the peptide, hydrolyzed protein, with zinc. And simple analytical method fot the chelation reaction was developed using MALDI-TOF mass spectrometry.

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SAFETY EVALUATION OF ADENOVIRUS-MEDIATED P16 GENE TRANSFER BY USING MICROARRAY AND 2D/MALDI-TOF

  • Park, Misun;Hoil Kang;Jaehee Pyo;Sinae Lim;Seungwan Jee;Miok Eom;Taikyung Ryeom;Kim, Okhee
    • Proceedings of the Korean Society of Toxicology Conference
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    • 2002.11b
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    • pp.196-196
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    • 2002
  • p16INK4a tumor suppressor gene transfer in the non-small cell lung cancer cells by transduction of recombinant adenovirus (Ad5CMV-p16) resulted in significant inhibition of cancer cell growth (Anticancer Res., 1998, 18:3257-3261). As a safety concern, we have investigated gene and protein expression after transduction of adenoviral vector (Ad5CMV-p16) in human non-small cell lung cancer (A549) cells by using microarray and 2D gel electrophoresis/ MALDI-TOF.(omitted)

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Ultrasonic Monitoring of Reaction of Fullerene$[C_{60}]$ with 3-Chloroperoxy Benzoic acid (풀러렌$[C_{60}]$과 3- Chloroperoxy Benzoic acid 반응의 초음파적 조사)

  • Ko, Weon-Bae;Buckin, Vitaly
    • Elastomers and Composites
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    • v.41 no.1
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    • pp.57-62
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    • 2006
  • The reaction of fullerene$[C_{60}]$ and 3-chloroperoxy benzoic acid in 1,2-dichlorobenzene was monitored by high resolution ultrasonic spectroscopy and the product of reaction by fullerene $[C_{60}]$ and 3-chloroperoxy benzoic acid in 1,2-dichlorobenzene was characterized using MALDI-TOF-MS spectra.

Rapid Identification of Vibrio Species Isolated from the Southern Coastal Regions of Korea by MALDI-TOF Mass Spectrometry and Comparison of MALDI Sample Preparation Methods

  • Cho, Youngjae;Kim, Eiseul;Han, Sun-Kyung;Yang, Seung-Min;Kim, Mi-ju;Kim, Hyun-Joong;Kim, Chang-Gyeom;Choo, Dong-Won;Kim, Young-Rok;Kim, Hae-Yeong
    • Journal of Microbiology and Biotechnology
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    • v.27 no.9
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    • pp.1593-1601
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    • 2017
  • Vibrio species are generally recognized as pathogens predominant in seafood along coastal areas. The food industry has sought to develop efficient microbial detection methods. Owing to the limits of conventional methods, this study aimed to establish a rapid identification method for Vibrio isolated from Korea, based on matrix-assisted laser-desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). Four different preparation procedures were compared to determine the appropriate means to pretreat Vibrio species, using 17 isolates and five reference strains. Extended direct transfer and full formic acid extraction methods using bacterial colonies on agar plates revealed very low identification rates. Formic acid and trifluoroacetic acid (TFA) extractions using bacterial broth cultures were also performed. All Vibrio isolates and reference strains prepared by TFA extraction were successfully identified to the species level (17/22, 77.3%) and to the genus level (5/22, 22.7%). Thus, TFA extraction was considered the most appropriate method to pretreat Vibrio species for MALDI-TOF MS. The remaining 33 isolates and two reference strains were prepared by TFA extraction and analyzed by MALDI-TOF MS. Overall, 50 isolates were identified to the species level (40/50, 80%) and to the genus level (10/50, 20%). All isolates were identified as 43 V. alginolyticus, six V. parahaemolyticus, and one V. vulnificus species. V. alginolyticus and V. parahaemolyticus were isolated from fish offal (87.5% and 12.5%, respectively), seawater (91.3%, 8.7%), and shellfish (62.5%, 37.5%), whereas V. alginolyticus and V. vulnificus were isolated from sediment (90.9% and 9.1%, respectively). This study established a reliable system of MALDI-TOF MS preparation and analysis for Vibrio identification.

Novel analysis procedure for red ginseng polysaccharides by matrix-assisted laser desorption/ionization time-of-flight/time-offlight mass spectrometry

  • Jin, Ye Rin;Oh, Myung Jin;Yuk, Heung Joo;An, Hyun Joo;Kim, Dong Seon
    • Journal of Ginseng Research
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    • v.45 no.5
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    • pp.539-545
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    • 2021
  • Background: Red ginseng polysaccharides (RGPs) have been acknowledged for their outstanding immunomodulation and anti-tumor activities. However, their studies are still limited by the complexity of their structural features, the absence of purification and enrichment methods, and the rarity of the analytical instruments that apply to the analysis of such macromolecules. Thus, this study is an attempt to establish a new mass spectrometry (MS)-based analysis procedure for RGPs. Methods: Saponin pre-excluded powder of RG (RG-SPEP, 10 mg) was treated with 200 µL of distilled water and centrifuged for 5 h at 1000 rpm and 85 ℃. Ethanol-based precipitation and centrifugation were applied to obtain RGPs from the heated extracts. Further, endo-carbohydrase treatments were performed to produce specific saccharide fragments. Solid-phase extraction (SPE) processes were implemented to purify and enrich the enzyme-treated RGPs, while matrix-assisted laser desorption/ionization time-of-flight/time-of-flight (MALDI-TOF/TOF) MS was employed for the partial structural analysis of the obtained RGPs. Results: Utilizing cellulase, porous graphitized carbon (PGC), hydrophilic interaction chromatography (HILIC), and MALDI-TOF/TOF MS, the neutral and acidic RGPs were qualitatively analyzed. Hexn and Hexn-18 (cellulose analogs) were determined to be novel neutral RGPs. Additionally, the [Unknown + Hexn] species were also determined as new acidic RGPs. Furthermore, HexAn (H) was determined as another form of the acidic RGPs. Conclusion: Compared to the previous methods of analysis, these unprecedented applications of HILIC-SPE and MALDI-TOF/TOF MS to analyze RGPs proved to be fairly effective for fractionating and detecting neutral and acidic components. This new procedure exhibits great potential as a specific tool for searching and determining various polysaccharides in many herbal medicines.

Effective Identification of Ochrobactrum anthropi Isolated from Clinical Specimens (임상검체에서 분리된 Ochrobactrum anthropi의 효과적인 동정)

  • Ko, Hyun-Mi;Jo, Jun-Hyeon;Baek, Hae-Gyeong
    • Korean Journal of Clinical Laboratory Science
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    • v.52 no.3
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    • pp.221-228
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    • 2020
  • Ochrobactrum anthropi is a non-fermentative oxidative gram-negative bacillus that produces oxidase. Distinguishing a mixed culture with non-fermenting bacteria having a similar appearance and oxidase-positive is difficult, and there is a limit to accurate identification with a biochemical identification system. This paper proposes that the Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry Platform (MALDI-TOF) method is useful for classifying bacteria that are difficult to identify using biochemical testing methods. As a result of analyzing five cases of O. anthropi examined using MicroScan, it took 6.5 days to the final report, which was 3.5 days more than the 3.0 days of E. coli. The pus sample in patient 5 was a mixed infection with Achromobacter xylosoxidans, and it took 11.3 days because of multiple subculture and retests. Four patients were over 60 years old with an underlying disease, and the possibility of opportunistic and nosocomial infections could not be excluded. Among them, samples collected after 92 days of hospitalization were resistant to imipenem and meropenem. Therefore, an examination using the MALDI-TOF method will be useful for the rapid and adequate treatment of patients with difficult identification, such as O. anthropi.

Proteomics 기법을 이용한 복제태반 분석

  • 김홍래;이혜란;강재구;윤종택;성한우;정진관;조민래;박창식;진동일
    • Proceedings of the KSAR Conference
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    • 2004.06a
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    • pp.238-238
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    • 2004
  • 세포 내에서 발현되고 있는 protein들의 양상을 분석하기 위한 기법으로 최근 proteomics에 기초하여 2차 전기영동과 MALDI-TOF MS에 의한 protein 분석방법이 개발되었는데, 특정 조직에 또는 특정 발생시기에 특이적으로 발현되는 protein의 발현양과 발현양상을 비교ㆍ분석하는데 매우 효과적으로 이용될 수 있다. 최근 체세포 핵이 식기술을 이용하여 동물의 복제가 성공하고 있지만, 임신 중이나 분만시 유사산이 많이 나타나 전반적인 효율이 크게 낮아 실용화에 지장을 초래하고 있다. (중략)

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Verification with of High Efficiency Chemical Binding System of a Physiologically Active Radioisotope Using ESI-TOF/Ms System (고효율의 ESI-TOF/Ms 시스템을 이용한 생리활성 항체와 방사성동위원소 표지용 착화제의 결합 검증)

  • Joh, Eun-Ha;Hong, Young Don;Choi, Sun Ju
    • YAKHAK HOEJI
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    • v.57 no.6
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    • pp.400-405
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    • 2013
  • In this study, we measured the complex efficiency of a physiologically active antibody, a chelator and radiosiotopes using the ESI-TOF/Ms system for develop good radiopharmaceuticals. For a precise measurement, TLC is a low accuracy method. Loading of same amount of sample is difficult for each test, and work to quantify accurately the results obtained through TLC cannot be afforded compared to the use of other analytical instruments. The method of analysis using a mass spectrometer is capable of a mass analysis of proteins for quantitative analysis. The conjugates of the chelator (CHX-A- DTPA) and the antibody (IgG) were separated through MWCO, and were analyzed using ESI-TOF and MALDI-TOF mass spectrometry. The analysis using MALDI-TOF is roughly divided into measurements on mass spectrometry. When conjugating a small molecular weight of CHX-A-DTPA and a large molecular weight of IgG, distinguishing the peak of the conjugate and the peak of IgG was difficult. However, an ESI-TOF mass spectrometer system is capable of an analysis of mass by decentralizing the IgG. It is utilized as a technique for measuring the metabolic processes during conjugation and the stability evaluation of radiopharmaceuticals. When establishing this technique, the accuracy of the overall radiophar-maceutical analysis is expected to be able to be improved.