Lead discovery for mammalian elongation of long chain fatty acids family 6 using a combination of high-throughput fluorescent-based assay and RapidFire mass spectrometry assay

Takamiya, Mari; Sakurai, Masaaki; Teranishi, Fumie; Ikeda, Tomoko; Kamiyama, Tsutomu; Asai, Akira

doi:10.1016/j.bbrc.2016.10.103

Cited by 9 publications

(3 citation statements)

References 15 publications

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“…Indeed, multiple examples of where RapidFire-MS was successfully employed to triage hits from light-based primary screens have been published and uniformly confirm the power of the approach to quickly eliminate false positives. These examples span a range of therapeutically relevant targets, including prostaglandin E synthase, 32 HIV-1 protease, 33 Elovl6, 34 and isocitrate dehydrogenase 1, 35 and highlight the ability of the RapidFire platform to reduce false-positive rates by as much as 83%. In a very comprehensive study, Adam et al applied RapidFire as a confirmatory technique to validate hits from primary screening campaigns against four different proteases.…”

Section: Rapidfire-msmentioning

confidence: 99%

High-Throughput Mass Spectrometry for Hit Identification: Current Landscape and Future Perspectives

et al. 2021

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Section: Rapidfire-msmentioning

confidence: 99%

High-Throughput Mass Spectrometry for Hit Identification: Current Landscape and Future Perspectives

et al. 2021

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“…High-throughput screening applying Lipinsky’s “rules of five” in structure–activity relationship studies combined with computational analyses and robotics is a frequently used methodology to identify the most promising compounds during drug discovery of low-molecular-weight targets in a fairly short period of time. The method also allows identification of specific positions in the low-molecular-weight targets for modifications to achieve desired properties, such as water − or fat solubility, − altered binding affinity by the modified strength of H-bonds, − and others.…”

Section: Introductionmentioning

confidence: 99%

Nanogel Catalysts for the Hydrolysis of Underivatized Disaccharides Identified by a Fast Screening Assay

Sharma

Striegler

2023

ACS Catal.

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Applying high-throughput synthetic and screening methods for the development of catalytic micro- and nanogels remains challenging due to the nature of the workflow. Typical procedures require the preparation of stable miniemulsions, followed by radical polymerization, purification of the obtained gels by extensive extraction or dialysis, and subsequent characterization of physical properties and examination of desired functions. To streamline the process, we altered the gel synthesis procedure and developed a screening protocol to identify gel compositions with the desired catalytic function. The assays identify five out of 50 synthesized gels from eight different cross-linkers with high potential to hydrolyze carbohydrates. A catalytic proficiency (k cat/K M × k non) of up to 1.4 × 106 and gel activity of 0.8 μmol h–1 mg–1 were observed placing the synthesized gels among the most proficient catalysts known for the hydrolysis of nonactivated glycosidic bonds.

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“…molecules from other biological samples have been analyzed in high throughput through online SPE cleanup using the Agilent RapidFire platform[72]. Acoustic droplet loading from microtiter plates via the OPP-ESI platform should also be applicable.…”

mentioning

confidence: 99%

Recent advances in high-throughput mass spectrometry that accelerates enzyme engineering for biofuel research

Zhang

2020

BMC Energy

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Enzymes play indispensable roles in producing biofuels, a sustainable and renewable source of transportation fuels. Lacking rational design rules, the development of industrially relevant enzyme catalysts relies heavily on highthroughput screening. However, few universal methods exist to rapidly characterize large-scale enzyme libraries. Therefore, assay development is necessary on an ad hoc basis to link enzyme properties to spectrophotometric signals and often requires the use of surrogate, optically active substrates. On the other hand, mass spectrometry (MS) performs label-free enzyme assays that utilize native substrates and is therefore generally applicable. But the analytical speed of MS is considered rate limiting, mainly due to the use of time-consuming chromatographic separation in traditional MS analysis. Thanks to new instrumentation and sample preparation methods, direct analyte introduction into a mass spectrometer without a prior chromatographic step can be achieved by laser, microfluidics, and acoustics, so that each sample can be analyzed within seconds. Here we review recent advances in MS platforms that improve the throughput of enzyme library screening and discuss how these advances can potentially facilitate biofuel research by providing high sensitivity, selectivity and quantitation that are difficult to obtain using traditional assays. We also highlight the limitations of current MS assays in studying biofuel-related enzymes and propose possible solutions.

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Lead discovery for mammalian elongation of long chain fatty acids family 6 using a combination of high-throughput fluorescent-based assay and RapidFire mass spectrometry assay

Cited by 9 publications

References 15 publications

High-Throughput Mass Spectrometry for Hit Identification: Current Landscape and Future Perspectives

High-Throughput Mass Spectrometry for Hit Identification: Current Landscape and Future Perspectives

Nanogel Catalysts for the Hydrolysis of Underivatized Disaccharides Identified by a Fast Screening Assay

Recent advances in high-throughput mass spectrometry that accelerates enzyme engineering for biofuel research

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