• 제목/요약/키워드: High-throughput Screening

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비선형 혼합효과모형에서의 로버스트 능형회귀 방법과 정량적 고속 대량 스크리닝 자료에의 응용 (Robust ridge regression for nonlinear mixed effects models with applications to quantitative high throughput screening assay data)

  • 유지선;임창원
    • 응용통계연구
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    • 제31권1호
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    • pp.123-137
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    • 2018
  • 비선형 혼합효과 모형은 다양한 분야에서 반복 측정 자료를 분석할 때 주로 사용된다. 비선형 혼합효과 모형은 개체 내 변동(intra-individual variation)에 대해 고려하는 제 1단계 개별수준모델(individual-level model)과 개체간 변동(inter-individual variation)에 대해 고려하는 제 2단계 개체군모델(population model)의 두 단계로 구성되어 있다. 비선형 혼합효과 모형의 첫 번째 단계인 개별수준모델은 비선형 회귀모형의 모수를 추정하는 것으로 일반적인 비선형 회귀모형과 같고, 주로 보통최소제곱추정 방법을 사용하여 모수를 추정한다. 그러나 최소제곱추정방법은 가정된 비선형 함수가 자료에 의해 명시적으로 드러나지 않는 경우 모수의 추정값과 그 표준오차가 극단적으로 커지는 문제가 발생할 수 있다. 본 논문에서는 최근에 비선형 회귀모형에서 제안된 능형회귀(ridge regression) 방법을 비선형 혼합효과 모형의 제 1단계 개별수준모델에 도입함으로써 이러한 문제를 해결할 수 있는 새로운 추정방법을 제안하였다. 제안된 추정량은 모의실험 연구를 통하여 기존의 표준적인 추정량과 그 성능을 비교하였다. 또한 미국의 National Toxicology Program으로부터 얻어진 정량적 대량고속 스크리닝(quantitative high throughput screening) 실제 자료를 사용하여 추정 방법들을 비교하였다.

Repeated Random Mutagenesis of ${\alpha}$-Amylase from Bacillus licheniformis for Improved pH Performance

  • Priyadharshini, Ramachandran;Manoharan, Shankar;Hemalatha, Devaraj;Gunasekaran, Paramasamy
    • Journal of Microbiology and Biotechnology
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    • 제20권12호
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    • pp.1696-1701
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    • 2010
  • The ${\alpha}$-amylases activity was improved by random mutagenesis and screening. A region comprising residues from the position 34-281 was randomly mutated in B. licheniformis ${\alpha}$-amylase (AmyL), and the library with mutations ranging from low, medium, and high frequencies was generated. The library was screened using an effective liquid-phase screening method to isolate mutants with an altered pH profile. The sequencing of improved variants indicated 2-5 amino acid changes. Among them, mutant TP8H5 showed an altered pH profile as compared with that of wild type. The sequencing of variant TP8H5 indicated 2 amino acid changes, Ile157Ser and Trp193Arg, which were located in the solvent accessible flexible loop region in domain B.

Evaluation of a New Episomal Vector Based on the GAP Promoter for Structural Genomics in Pichia pastoris

  • Hong In-Pyo;Anderson Stephen;Choi Shin-Geon
    • Journal of Microbiology and Biotechnology
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    • 제16권9호
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    • pp.1362-1368
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    • 2006
  • A new constitutive episomal expression vector, pGAPZ-E, was constructed and used for initial screening of eukaryotic target gene expression in Pichia pastoris. Two reporter genes such as beta-galactosidase gene and GFPuv gene were overexpressed in P. pastoris. The expression level of the episomal pGAPZ-E strain was higher than that of the integrated form when the beta-galactosidase gene was used as the reporter gene in P. pastoris X33. The avoiding of both the integration procedure and an induction step simplified the overall screening process for eukaryotic target gene expression in P. pastoris. Nine human protein targets from the Core 50, family of Northeast Structural Genomics Consortium (http://www.nesg.org), which were intractable when expressed in E. coli, were subjected to rapid screening for soluble expression in P. pastoris. HR547, HR919, and HR1697 human proteins, which had previously been found to express poorly or to be insoluble in E. coli, expressed in soluble form in P. pastoris. Therefore, the new episomal GAP promoter vector provides a convenient and alternative system for high-throughput screening of eukaryotic protein expression in P. pastoris.

High-Throughput In Vitro Screening of Changed Algal Community Structure Using the PhotoBiobox

  • Cho, Dae-Hyun;Cho, Kichul;Heo, Jina;Kim, Urim;Lee, Yong Jae;Choi, Dong-Yun;Yoo, Chan;Kim, Hee-Sik;Bae, Seunghee
    • Journal of Microbiology and Biotechnology
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    • 제30권11호
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    • pp.1785-1791
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    • 2020
  • In a previous study, the sequential optimization and regulation of environmental parameters using the PhotoBiobox were demonstrated with high-throughput screening tests. In this study, we estimated changes in the biovolume-based composition of a polyculture built in vitro and composed of three algal strains: Chlorella sp., Scenedesmus sp., and Parachlorella sp. We performed this work using the PhotoBiobox under different temperatures (10-36℃) and light intensities (50-700 μmol m-2 s-1) in air and in 5% CO2. In 5% CO2, Chlorella sp. exhibited better adaptation to high temperatures than in air conditions. Pearson's correlation analysis showed that the composition of Parachlorella sp. was highly related to temperature whereas Chlorella sp. and Scenedesmus sp. showed negative correlations in both air and 5% CO2. Furthermore, light intensity slightly affected the composition of Scenedesmus sp., whereas no significant effect was observed in other species. Based on these results, it is speculated that temperature is an important factor in influencing changes in algal polyculture community structure (PCS). These results further confirm that the PhotoBiobox is a convenient and available tool for performance of lab-scale experiments on PCS changes. The application of the PhotoBiobox in PCS studies will provide new insight into polyculture-based ecology.

Design and Combonatorial Synthesis of Selenoflavonoids and Indole Compounds for Neuroprotective Drug

  • Jeong, Jin-Hyun
    • 대한약학회:학술대회논문집
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    • 대한약학회 2003년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.1
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    • pp.69-71
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    • 2003
  • As recent as 10 years ago, a method of developing new medicine was developed by a new compounding method moving away from an existing one. Combinatorial chemistry made it easier to combine various kinds of compounds in a very short time and with little effort from existing methods. Through combinatorial chemistry, a number of compounds were synthesized using HTS(High Throughput Screening), with many reports reaching a clinical stage in search of new candidate material. (omitted)

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Screening of novel alkaloid inhibitors for vascular endothelial growth factor in cancer cells: an integrated computational approach

  • Shahik, Shah Md.;Salauddin, Asma;Hossain, Md. Shakhawat;Noyon, Sajjad Hossain;Moin, Abu Tayab;Mizan, Shagufta;Raza, Md. Thosif
    • Genomics & Informatics
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    • 제19권1호
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    • pp.6.1-6.10
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    • 2021
  • Vascular endothelial growth factor (VEGF) is expressed at elevated levels by most cancer cells, which can stimulate vascular endothelial cell growth, survival, proliferation as well as trigger angiogenesis modulated by VEGF and VEGFR (a tyrosine kinase receptor) signaling. The angiogenic effects of the VEGF family are thought to be primarily mediated through the interaction of VEGF with VEGFR-2. Targeting this signaling molecule and its receptor is a novel approach for blocking angiogenesis. In recent years virtual high throughput screening has emerged as a widely accepted powerful technique in the identification of novel and diverse leads. The high resolution X-ray structure of VEGF has paved the way to introduce new small molecular inhibitors by structure-based virtual screening. In this study using different alkaloid molecules as potential novel inhibitors of VEGF, we proposed three alkaloid candidates for inhibiting VEGF and VEGFR mediated angiogenesis. As these three alkaloid compounds exhibited high scoring functions, which also highlights their high binding ability, it is evident that these alkaloids can be taken to further drug development pipelines for use as novel lead compounds to design new and effective drugs against cancer.

Tyrosine Phenol-Lyase의 고속탐색기술 개발 및 방향성 분자진화 (High Throughput Screening and Directed Evolution of Tyrosine Phenol-Lyase)

  • 최수림;나유진;김도영;송재준;홍승표;성문희;이승구
    • 한국미생물·생명공학회지
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    • 제34권1호
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    • pp.58-62
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    • 2006
  • 티로신 및 방향족 아미노산 유도체의 생물학적 합성에 이용되는 유용 효소인 tyrosine phenol-lyase(TPL) 활성을 평판배지에서 정량적으로 감지할 수 있는 기술을 개발하였다. 불투명 평판배지(turbid plate)의 제조과정은 먼저, 2 N염산과 DMSO로 조성된 공용매에 난용성 티로신을 400 mM 농도로 녹여서 배지에 가하고, 온도를 조절하여 니들형 티로신을 형성하도록 결정화를 유도하는 것이었다. 니들형 티로신은 일반적인 컬럼형에 비하여 약 $5{\sim}6$배 낮은 농도인 3.6g/L에서도 불투명 평판배지를 제조할 수 있었으며, 효소활성에 의하여 쉽게 분해되고, TPL활성의 고감도 고속탐색기술 개발에 적합하였다. 이 불투명 평판배지에 변이유발PCR법으로 제조한 TPL 라이브러리를 도말하고, 단일 콜로니 주변에서 형성되는 투명환의 크기와 실측한 TPL활성을 비교한 결과 직접적인 비례관계가 있음을 확인하였다. 따라서 난용성 물질의 미세입자를 평판배지에서 직접 발생시킨 후, 효소활성에 의한 투명환의 형성을 정량 관찰하는 본 연구의 방법은 신규 고활성 TPL의 분리 및 방향성 분자진화 등을 위한 고속탐색기술에 유용하게 사용될 수 있다.