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98.127.152.74: /* Recent advances */ added an advancement with a reduction in time from 24 hours to less than 2 and allows for higher drug loadings previously not possible and citation.

2024-11-13T18:49:08Z

Recent advances: added an advancement with a reduction in time from 24 hours to less than 2 and allows for higher drug loadings previously not possible and citation.

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← Previous revision		Revision as of 03:58, 5 November 2025
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	{{Short description\|Drug discovery technique}}		{{Short description\|Drug discovery technique}}
	[[File:Chemical Genomics Robot.jpg\|thumb\|300px\|High-throughput screening robots]]		[[File:Chemical Genomics Robot.jpg\|thumb\|300px\|High-throughput screening robots]]
	'''High-throughput screening''' ('''HTS''') is a method for scientific discovery especially used in [[drug discovery]] and relevant to the fields of [[biology]], [[materials science]]<ref>{{cite journal \|last1=Zhao \|first1=Yicheng \|last2=Heumueller \|first2=Thomas \|last3=Zhang \|first3=Jiyun \|last4=Luo \|first4=Junsheng \|last5=Kasian \|first5=Olga \|last6=Langner \|first6=Stefan \|last7=Kupfer \|first7=Christian \|last8=Liu \|first8=Bowen \|last9=Zhong \|first9=Yu \|last10=Elia \|first10=Jack \|last11=Osvet \|first11=Andres \|last12=Wu \|first12=Jianchang \|last13=Liu \|first13=Chao \|last14=Wan \|first14=Zhongquan \|last15=Jia \|first15=Chunyang \|last16=Li \|first16=Ning \|last17=Hauch \|first17=Jens \|last18=Brabec \|first18=Christoph J. \|title=A bilayer conducting polymer structure for planar perovskite solar cells with over 1,400 hours operational stability at elevated temperatures \|journal=Nature Energy \|date=16 December 2021 \|volume=7 \|issue=2 \|pages=144–152 \|doi=10.1038/s41560-021-00953-z \|bibcode=2022NatEn...7..144Z \|s2cid=245285868 \|language=en \|issn=2058-7546}}</ref> and [[chemistry]].<ref>Inglese J and Auld DS. (2009) Application of High Throughput Screening (HTS) Techniques: Applications in Chemical Biology in Wiley Encyclopedia of Chemical Biology (Wiley & Sons, Inc., Hoboken, NJ) Vol 2, pp 260–274 doi/10.1002/9780470048672.wecb223.</ref><ref>{{cite journal \| title = Impact of high-throughput screening in biomedical research \|author1 = Macarron, R. \|author2 = Banks, M.N.\|author3 = Bojanic, D.\| author4 = Burns, D.J.\|author5 = Cirovic, D.A.\|author6 = Garyantes, T.\|author7 = Green, D.V.\| author8 = Hertzberg, R.P. \|author9 = Janzen, W.P.\|author10 = Paslay, J.W.\|author11 = Schopfer, U.\| author12 = Sittampalam, G.S. \| journal = Nat Rev Drug Discov\|date = 2011 \|volume = 10\|issue = 3 \|pages = 188–195\|pmid = 21358738 \| doi = 10.1038/nrd3368\|s2cid = 205477370 }}</ref> Using [[robotics]], data processing/control software, liquid handling devices, and sensitive detectors, high-throughput screening allows a researcher to quickly conduct millions of chemical, genetic, or pharmacological tests. Through this process one can quickly recognize active compounds, antibodies, or genes that modulate a particular biomolecular pathway. The results of these experiments provide starting points for drug design and for understanding the noninteraction or role of a particular location.		'''High-throughput screening''' ('''HTS''') is a method for scientific discovery especially used in [[drug discovery]] and relevant to the fields of [[biology]], [[materials science]]<ref>{{cite journal \|last1=Zhao \|first1=Yicheng \|last2=Heumueller \|first2=Thomas \|last3=Zhang \|first3=Jiyun \|last4=Luo \|first4=Junsheng \|last5=Kasian \|first5=Olga \|last6=Langner \|first6=Stefan \|last7=Kupfer \|first7=Christian \|last8=Liu \|first8=Bowen \|last9=Zhong \|first9=Yu \|last10=Elia \|first10=Jack \|last11=Osvet \|first11=Andres \|last12=Wu \|first12=Jianchang \|last13=Liu \|first13=Chao \|last14=Wan \|first14=Zhongquan \|last15=Jia \|first15=Chunyang \|last16=Li \|first16=Ning \|last17=Hauch \|first17=Jens \|last18=Brabec \|first18=Christoph J. \|title=A bilayer conducting polymer structure for planar perovskite solar cells with over 1,400 hours operational stability at elevated temperatures \|journal=Nature Energy \|date=16 December 2021 \|volume=7 \|issue=2 \|pages=144–152 \|doi=10.1038/s41560-021-00953-z \|bibcode=2022NatEn...7..144Z \|s2cid=245285868 \|language=en \|issn=2058-7546}}</ref> and [[chemistry]].<ref>Inglese J and Auld DS. (2009) Application of High Throughput Screening (HTS) Techniques: Applications in Chemical Biology in Wiley Encyclopedia of Chemical Biology (Wiley & Sons, Inc., Hoboken, NJ) Vol 2, pp 260–274 doi/10.1002/9780470048672.wecb223.</ref><ref>{{cite journal \| title = Impact of high-throughput screening in biomedical research \|author1 = Macarron, R. \|author2 = Banks, M.N.\|author3 = Bojanic, D.\| author4 = Burns, D.J.\|author5 = Cirovic, D.A.\|author6 = Garyantes, T.\|author7 = Green, D.V.\| author8 = Hertzberg, R.P. \|author9 = Janzen, W.P.\|author10 = Paslay, J.W.\|author11 = Schopfer, U.\| author12 = Sittampalam, G.S. \| journal = Nat Rev Drug Discov\|date = 2011 \|volume = 10\|issue = 3 \|pages = 188–195\|pmid = 21358738 \| doi = 10.1038/nrd3368\|s2cid = 205477370 }}</ref> Using [[robotics]], data processing/control software, liquid handling devices, and sensitive detectors, high-throughput screening allows a researcher to quickly conduct millions of chemical, genetic, or pharmacological tests. Through this process one can quickly recognize active compounds, antibodies, or genes that modulate a particular biomolecular pathway. The results of these experiments provide starting points for [[drug design]] and for understanding the noninteraction or role of a particular location.

	==Assay plate preparation==		==Assay plate preparation==
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	==Automation systems==		==Automation systems==
	[[File:Assay plate carousel.jpg\|thumb\|A carousel system to store assay plates for high storage capacity and high speed access]]		[[File:Assay plate carousel.jpg\|thumb\|A carousel system to store assay plates for high storage capacity and high speed access]]
	[[Automation]] is an essential element in HTS's usefulness. Typically, an integrated [[robot]] system consisting of one or more robots transports assay-microplates from station to station for sample and reagent addition, mixing, incubation, and finally readout or detection. An HTS system can usually prepare, incubate, and analyze many plates simultaneously, further speeding the data-collection process. HTS robots that can test up to 100,000 compounds per day currently exist.<ref name="pmid15183323">{{cite journal \|vauthors=Hann MM, Oprea TI \|title=Pursuing the leadlikeness concept in pharmaceutical research \|journal=Curr Opin Chem Biol \|volume=8 \|issue=3 \|pages=255–63 \|date=June 2004 \|pmid=15183323 \|doi=10.1016/j.cbpa.2004.04.003 \|url=https://zenodo.org/record/896094}}</ref><ref name="caraus974">{{Cite journal\|last1=Caraus\|first1=I.\|last2=Alsuwailem\|first2=A. A.\|last3=Nadon\|first3=R.\|last4=Makarenkov\|first4=V.\|date=2015-11-01\|title=Detecting and overcoming systematic bias in high-throughput screening technologies: a comprehensive review of practical issues and methodological solutions\|url=https://academic.oup.com/bib/article/16/6/974/225604/Detecting-and-overcoming-systematic-bias-in-high\|journal=Briefings in Bioinformatics\|language=en\|volume=16\|issue=6\|pages=974–986\|doi=10.1093/bib/bbv004\|pmid=25750417\|issn=1467-5463\|doi-access=free}}</ref> [[Colony Picker\|Automatic colony pickers]] pick thousands of microbial colonies for high throughput genetic screening.<ref name="HeddleMazaleyrat2007">{{cite journal\|last1=Heddle\|first1=C.\|last2=Mazaleyrat\|first2=S. L.\|title=Development of a screening platform for directed evolution using the reef coral fluorescent protein ZsGreen as a solubility reporter\|journal=Protein Engineering Design and Selection\|volume=20\|issue=7\|year=2007\|pages=327–337\|issn=1741-0126\|doi=10.1093/protein/gzm024\|pmid=17584755\|doi-access=free}}</ref> The term uHTS or ''ultra-high-throughput screening'' refers (circa 2008) to screening in excess of 100,000 compounds per day.<ref name="MichaelAuld2008">{{cite journal\|last1=Michael\|first1=Sam\|last2=Auld\|first2=Douglas\|last3=Klumpp\|first3=Carleen\|last4=Jadhav\|first4=Ajit\|last5=Zheng\|first5=Wei\|last6=Thorne\|first6=Natasha\|last7=Austin\|first7=Christopher P.\|last8=Inglese\|first8=James\|last9=Simeonov\|first9=Anton\|title=A Robotic Platform for Quantitative High-Throughput Screening\|journal=ASSAY and Drug Development Technologies\|volume=6\|issue=5\|year=2008\|pages=637–657\|issn=1540-658X\|doi=10.1089/adt.2008.150\|pmid=19035846\|pmc=2651822}}</ref>		[[Automation]] is an essential element in HTS's usefulness. Typically, an integrated [[robot]] system consisting of one or more robots transports assay-microplates from station to station for sample and [[reagent]] addition, mixing, incubation, and finally readout or detection. An HTS system can usually prepare, incubate, and analyze many plates simultaneously, further speeding the data-collection process. HTS robots that can test up to 100,000 compounds per day currently exist.<ref name="pmid15183323">{{cite journal \|vauthors=Hann MM, Oprea TI \|title=Pursuing the leadlikeness concept in pharmaceutical research \|journal=Curr Opin Chem Biol \|volume=8 \|issue=3 \|pages=255–63 \|date=June 2004 \|pmid=15183323 \|doi=10.1016/j.cbpa.2004.04.003 \|url=https://zenodo.org/record/896094}}</ref><ref name="caraus974">{{Cite journal\|last1=Caraus\|first1=I.\|last2=Alsuwailem\|first2=A. A.\|last3=Nadon\|first3=R.\|last4=Makarenkov\|first4=V.\|date=2015-11-01\|title=Detecting and overcoming systematic bias in high-throughput screening technologies: a comprehensive review of practical issues and methodological solutions\|url=https://academic.oup.com/bib/article/16/6/974/225604/Detecting-and-overcoming-systematic-bias-in-high\|journal=Briefings in Bioinformatics\|language=en\|volume=16\|issue=6\|pages=974–986\|doi=10.1093/bib/bbv004\|pmid=25750417\|issn=1467-5463\|doi-access=free}}</ref> [[Colony Picker\|Automatic colony pickers]] pick thousands of microbial colonies for high throughput genetic screening.<ref name="HeddleMazaleyrat2007">{{cite journal\|last1=Heddle\|first1=C.\|last2=Mazaleyrat\|first2=S. L.\|title=Development of a screening platform for directed evolution using the reef coral fluorescent protein ZsGreen as a solubility reporter\|journal=Protein Engineering Design and Selection\|volume=20\|issue=7\|year=2007\|pages=327–337\|issn=1741-0126\|doi=10.1093/protein/gzm024\|pmid=17584755\|doi-access=free}}</ref> The term uHTS or ''ultra-high-throughput screening'' refers (circa 2008) to screening in excess of 100,000 compounds per day.<ref name="MichaelAuld2008">{{cite journal\|last1=Michael\|first1=Sam\|last2=Auld\|first2=Douglas\|last3=Klumpp\|first3=Carleen\|last4=Jadhav\|first4=Ajit\|last5=Zheng\|first5=Wei\|last6=Thorne\|first6=Natasha\|last7=Austin\|first7=Christopher P.\|last8=Inglese\|first8=James\|last9=Simeonov\|first9=Anton\|title=A Robotic Platform for Quantitative High-Throughput Screening\|journal=ASSAY and Drug Development Technologies\|volume=6\|issue=5\|year=2008\|pages=637–657\|issn=1540-658X\|doi=10.1089/adt.2008.150\|pmid=19035846\|pmc=2651822}}</ref>

	== Experimental design and data analysis ==		== Experimental design and data analysis ==
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	=== Quality control ===		=== Quality control ===
	High-quality HTS assays are critical in HTS experiments. The development of high-quality HTS assays requires the integration of both experimental and computational approaches for quality control (QC). Three important means of QC are (i) good plate design, (ii) the selection of effective positive and negative chemical/biological controls, and (iii) the development of effective QC metrics to measure the degree of differentiation so that assays with inferior data quality can be identified.		High-quality HTS assays are critical in HTS experiments. The development of high-quality HTS assays requires the integration of both experimental and computational approaches for quality control (QC). Three important means of QC are (i) good plate design, (ii) the selection of effective positive and negative chemical/biological controls, and (iii) the development of effective QC metrics to measure the degree of differentiation so that assays with inferior [[data quality]] can be identified.
	<ref name=ZhangetalJBS2008>{{cite journal \|vauthors=Zhang XH, Espeseth AS, Johnson EN, Chin J, Gates A, Mitnaul LJ, Marine SD, Tian J, Stec EM, Kunapuli P, Holder DJ, Heyse JF, Strulocivi B, Ferrer M \|title=Integrating experimental and analytic approaches to improve data quality in genome-scale RNAi screens		<ref name=ZhangetalJBS2008>{{cite journal \|vauthors=Zhang XH, Espeseth AS, Johnson EN, Chin J, Gates A, Mitnaul LJ, Marine SD, Tian J, Stec EM, Kunapuli P, Holder DJ, Heyse JF, Strulocivi B, Ferrer M \|title=Integrating experimental and analytic approaches to improve data quality in genome-scale RNAi screens
	\|journal=Journal of Biomolecular Screening \|volume=13 \|issue= 5\|pages=378–89		\|journal=Journal of Biomolecular Screening \|volume=13 \|issue= 5\|pages=378–89
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	In 2013, researchers have disclosed an approach with small molecules from plants. In general, it is essential to provide high-quality proof-of-concept validations early in the drug discovery process. Here technologies that enable the identification of potent, selective, and bioavailable chemical probes are of crucial interest, even if the resulting compounds require further optimization for development into a pharmaceutical product. Nuclear receptor RORα, a protein that has been targeted for more than a decade to identify potent and bioavailable agonists, was used as an example of a very challenging drug target. Hits are confirmed at the screening step due to the bell-shaped curve. This method is very similar to the quantitative HTS method (screening and hit confirmation at the same time), except that using this approach greatly decreases the data point number and can screen easily more than 100.000 biological relevant compounds.<ref>Helleboid S, Haug C, Lamottke K, et al. The Identification of Naturally Occurring Neoruscogenin as a Bioavailable, Potent, and High-Affinity Agonist of the Nuclear Receptor RORα (NR1F1). Journal of Biomolecular Screening. 2014;19(3):399-406. https://doi.org/10.1177/1087057113497095.</ref>		In 2013, researchers have disclosed an approach with small molecules from plants. In general, it is essential to provide high-quality proof-of-concept validations early in the drug discovery process. Here technologies that enable the identification of potent, selective, and bioavailable chemical probes are of crucial interest, even if the resulting compounds require further optimization for development into a pharmaceutical product. Nuclear receptor RORα, a protein that has been targeted for more than a decade to identify potent and bioavailable agonists, was used as an example of a very challenging drug target. Hits are confirmed at the screening step due to the bell-shaped curve. This method is very similar to the quantitative HTS method (screening and hit confirmation at the same time), except that using this approach greatly decreases the data point number and can screen easily more than 100.000 biological relevant compounds.<ref>Helleboid S, Haug C, Lamottke K, et al. The Identification of Naturally Occurring Neoruscogenin as a Bioavailable, Potent, and High-Affinity Agonist of the Nuclear Receptor RORα (NR1F1). Journal of Biomolecular Screening. 2014;19(3):399-406. https://doi.org/10.1177/1087057113497095.</ref>

	Switching from an orbital shaker, which required milling times of 24 hours and at least 10 mg of drug compound to a ResonantAcoustic mixer, Merck reported reduced processing time to less than 2 hours on only 1-2 mg of drug compound per well. Merck also indicated the acoustic milling approach allows for the preparation of high dose nanosuspension formulations that could not be obtained using conventional milling equipment.<ref name="a501">{{cite journal \| ~~last~~=Leung \| ~~first~~=Dennis H. \| last2=Lamberto \| first2=David J. \| last3=Liu \| first3=Lina \| last4=Kwong \| first4=Elizabeth \| last5=Nelson \| first5=Todd \| last6=Rhodes \| first6=Timothy \| last7=Bak \| first7=Annette \| title=A new and improved method for the preparation of drug nanosuspension formulations using acoustic mixing technology \| journal=International Journal of Pharmaceutics \| publisher=Elsevier BV \| volume=473 \| issue=~~1-2~~ \| year=2014 \| doi=10.1016/j.ijpharm.2014.05.003 \| pages=10–19}}</ref>		Switching from an orbital shaker, which required milling times of 24 hours and at least 10 mg of drug compound to a ResonantAcoustic mixer, Merck reported reduced processing time to less than 2 hours on only 1-2 mg of drug compound per well. Merck also indicated the acoustic milling approach allows for the preparation of high dose nanosuspension formulations that could not be obtained using conventional milling equipment.<ref name="a501">{{cite journal \| last1=Leung \| first1=Dennis H. \| last2=Lamberto \| first2=David J. \| last3=Liu \| first3=Lina \| last4=Kwong \| first4=Elizabeth \| last5=Nelson \| first5=Todd \| last6=Rhodes \| first6=Timothy \| last7=Bak \| first7=Annette \| title=A new and improved method for the preparation of drug nanosuspension formulations using acoustic mixing technology \| journal=International Journal of Pharmaceutics \| publisher=Elsevier BV \| volume=473 \| issue=1–2 \| year=2014 \| doi=10.1016/j.ijpharm.2014.05.003 \| pages=10–19 \| pmid=24984068 }}</ref>

	Whereby traditional HTS drug discovery uses purified proteins or intact cells, recent development of the technology is associated with the use of intact living organisms, like the nematode ''[[Caenorhabditis elegans]]'' and zebrafish ([[Zebrafish\|''Danio rerio'']]).<ref>{{cite journal \|vauthors=Atanasov AG, Waltenberger B, Pferschy-Wenzig EM, Linder T, Wawrosch C, Uhrin P, Temml V, Wang L, Schwaiger S, Heiss EH, Rollinger JM, Schuster D, Breuss JM, Bochkov V, Mihovilovic MD, Kopp B, Bauer R, Dirsch VM, Stuppner H \| year = 2015 \| title = Discovery and resupply of pharmacologically active plant-derived natural products: A review \| doi = 10.1016/j.biotechadv.2015.08.001 \| journal = Biotechnol. Adv. \| volume = 33\| issue = 8\| pages = 1582–614\| pmid = 26281720 \| pmc=4748402}}</ref>		Whereby traditional HTS drug discovery uses purified proteins or intact cells, recent development of the technology is associated with the use of intact living organisms, like the nematode ''[[Caenorhabditis elegans]]'' and zebrafish ([[Zebrafish\|''Danio rerio'']]).<ref>{{cite journal \|vauthors=Atanasov AG, Waltenberger B, Pferschy-Wenzig EM, Linder T, Wawrosch C, Uhrin P, Temml V, Wang L, Schwaiger S, Heiss EH, Rollinger JM, Schuster D, Breuss JM, Bochkov V, Mihovilovic MD, Kopp B, Bauer R, Dirsch VM, Stuppner H \| year = 2015 \| title = Discovery and resupply of pharmacologically active plant-derived natural products: A review \| doi = 10.1016/j.biotechadv.2015.08.001 \| journal = Biotechnol. Adv. \| volume = 33\| issue = 8\| pages = 1582–614\| pmid = 26281720 \| pmc=4748402}}</ref>
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	==External links==		==External links==
	* [~~http~~://sen.sourceforge.net/ Open Screening Environment]		* [https://sen.sourceforge.net/ Open Screening Environment]
	* [https://web.archive.org/web/20101214123953/http://labmanager.com/articles.asp?ID=481 Setting up High-Throughput Screening Laboratory] (Koppal, Lab Manager Magazine)		* [https://web.archive.org/web/20101214123953/http://labmanager.com/articles.asp?ID=481 Setting up High-Throughput Screening Laboratory] (Koppal, Lab Manager Magazine)
	* [https://wayback.archive-it.org/all/20121010200014/http://spotlite.nih.gov/assay/index.php/Table_of_Contents Assay Guidance Manual] (NIH, NCGC)		* [https://wayback.archive-it.org/all/20121010200014/http://spotlite.nih.gov/assay/index.php/Table_of_Contents Assay Guidance Manual] (NIH, NCGC)

High-throughput screening - Revision history

imported>VineWeaver: /* growthexperiments-addlink-summary-summary:3|0|0 */

98.127.152.74: /* Recent advances */ added an advancement with a reduction in time from 24 hours to less than 2 and allows for higher drug loadings previously not possible and citation.