Applications & Publications
Technical Notes
Encapsulation in monodispersed hydrogel microspheres enables fast and sensitive phenotypic analyses using COPAS large particle flow cytometry. (QTN-019)
March 01, 2010
Publications
IgE Epitope Profiling for Allergy Diagnosis and Therapy – Parallel Analysis of a Multitude of Potential Linear Epitopes Using a High Throughput Screening Platform
Krause et al. September 30, 2020 Front. Immunol., 30 September 2020 | https://doi.org/10.3389/fimmu.2020.565243
View AbstractIgE Epitope Profiling for Allergy Diagnosis and Therapy – Parallel Analysis of a Multitude of Potential Linear Epitopes Using a High Throughput Screening Platform
Many objects are too large and too fragile for conventional flowcytometry. Manual microscopic manipulation of model organisms is tedious, subjective, and limits the size and scope of experiments. Similarly, “bead pickers”offer only one-at-a-time manipulation of combinatorial libraries.
Instruments are now available to automate this process of sorting large (40-1,000 micron) particles in a continuously flowing stream at a rate of 10-30 objects/second. Using object size, optical density, and intensity of fluorescent markers as sorting criteria, selected beads or animals can be dispensed in multi-well plates for further validation of hits. A gentle pneumatic sorting mechanismlocated after the flow cell avoids harming or changing sensitive objects, thereby making the instrument suitable for live biological materials or sensitive chemistries. Multiple fluorescence excitation and emission wavelengths are available, with sensitivity for GFP, YFP,DsRed, as well as the commercially availablefluorophorescommonly used incombi-chem.
Beads can be quickly screened before the assay step to eliminatecomplications due to broken beads, bead aggregates or beads with auto-fluorescence. Binding assays, for example, can be shrunk to a single-bead, thereby minimizing the amount of precious target proteins/peptides and other expensive reagents required for a screen. Use of high-capacity 50-500 micron beads leaves sufficient residual chemistry on a single bead for post factoNMR and MS structure determinations.
Today, automation, increased sensitivity and quantitative measurements, enable larger / faster screens of both model organisms and combinatorial libraries. Several examples will be discussed.
Identification and Characterisation of a pH-stable GFP
Tania M. Roberts, Fabian Rudolf, Andreas Meyer, Rene Pellaux, Ellis Whitehead, Sven Panke, and Martin Held June 21, 2016 Scientific Reports, (2016) London: Nature Publishing Group.
Identification and Characterisation of a pH-stable GFP
High throughput screening of one-bead-one-compound peptide libraries using intact cells
Choi-Fong Cho, Babak Behnam Azad, Leonard G. Luyt, and John Lewis July 02, 2013 ACS Comb. Sci., Just Accepted Manuscript • DOI: 10.1021/co4000584 • Publication Date (Web): 02 Jul 2013
High throughput screening of one-bead-one-compound peptide libraries using intact cells
Flow-based pipeline for systematic modulation and analysis of 3D tumor microenvironments.
Li CY, Wood DK, Huang JH, Bhatia SN.
Chemical Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, United States.
April 23, 2013
Lab Chip. 2013 Apr 23;13(10):1969-78. doi: 10.1039/c3lc41300d. Epub 2013 Apr 5.
Flow-based pipeline for systematic modulation and analysis of 3D tumor microenvironments.
A two-channel detection method for autofluorescence correction and efficient on-bead screening of one-bead one-compound combinatorial libraries using the COPAS fluorescence activated bead sorting system
Martin Hintersteiner¹ and Manfred Auer² March 12, 2013 2013 Methods Appl. Fluoresc. 1 017001 doi:10.1088/2050-6120/1/1/017001
A two-channel detection method for autofluorescence correction and efficient on-bead screening of one-bead one-compound combinatorial libraries using the COPAS fluorescence activated bead sorting system
Process Automation toward Ultra-High-Throughput Screening of Combinatorial One-Bead-One-Compound (OBOC) Peptide Libraries
Junhoe Cha, Jaehong Lim, Yiran Zheng, Sylvia Tan, Yi Li Ang, Jessica Oon, Mei Wei Ang, Jingjing Ling, Marcus Bode, Su Seong Lee June 17, 2012 Journal of Laboratory Automation 2012 17: 186 originally published online 14 February 2012 Institute of Bioengineering and Nanotechnology, Singapore.
Process Automation toward Ultra-High-Throughput Screening of Combinatorial One-Bead-One-Compound (OBOC) Peptide Libraries
TFIIS is required for the balanced expression of the genes encoding ribosomal components under transcriptional stress
Fernando Gómez-Herreros, Lola de Miguel-Jiménez, Macarena Morillo-Huesca, Lidia Delgado-Ramos, María C. Muñoz-Centeno and Sebastián Chávez April 27, 2012 Nucleic Acids Research, 2012, 1–12
View AbstractTFIIS is required for the balanced expression of the genes encoding ribosomal components under transcriptional stress
Microbial encapsulation in monodisperse hydrogel microspheres enables fast and sensitive phenotypic analyses using flow cytometers
Lidia Delgado¹, Gloria Jurado², Gema Galayo², Elena Ogalla², Lourdes Moreno³, Juan C Rodríguez-Aguilera4, Ángel Cebolla5, Carolina Sousa³, María Flores² and Sebastián Chávez¹ February 28, 2011 Conference Presentation at: La Société Française de Microbiologie et l’Association Française de Cytométrie 1) Department of Genetics, Universidad de Sevilla, Seville, Spain. 2) Ingeniatrics Tecnologías SL, Spain. 3) Department of Microbiology and Parasitology, Universidad de Sevilla, Seville, Spain. 4) Centro Andaluz de Biología del Desarrollo, Universidad Pablo de Olavide, Seville, Spain. 5) Biomedal SL, Spain
Microbial encapsulation in monodisperse hydrogel microspheres enables fast and sensitive phenotypic analyses using flow cytometers
Multiplexed, high-throughput analysis of 3D microtissue suspensions.
Chen AA, Underhill GH, Bhatia SN October 04, 2010 Integr Biol (Camb). 2010 Oct 4;2(10):517-27. Epub 2010 Sep 1. Massachusetts Institute of Technology, Cambridge, 02139, USA.
View AbstractMultiplexed, high-throughput analysis of 3D microtissue suspensions.
Three-dimensional (3D) tissue models have significantly improved our understanding of structure/function relationships and promise to lead to new advances in regenerative medicine. However, despite the expanding diversity of 3D tissue fabrication methods, approaches for functional assessment have been relatively limited. Here, we describe the fabrication of microtissue (μ-tissue) suspensions and their quantitative evaluation with techniques capable of analyzing large sample numbers and performing multiplexed parallel analysis. We applied this platform to 3D μ-tissues representing multiple stages of liver development and disease including: embryonic stem cells, bipotential hepatic progenitors, mature hepatocytes, and hepatoma cells photoencapsulated in polyethylene glycol hydrogels. Multiparametric μ-tissue cytometry enabled quantitation of fluorescent reporter expression within populations of intact μ-tissues (n≥ 10²-10³) and sorting-based enrichment of subsets for subsequent studies. Further, 3D μ-tissues could be implanted in vivo, respond to systemic stimuli, retrieved and quantitatively assessed. In order to facilitate multiplexed 'pooled' experimentation, fluorescent labeling strategies were developed and utilized to investigate the impact of μ-tissue composition and exposure to soluble factors. In particular, examination of drug/gene interactions on collections of 3D hepatoma μ-tissues indicated synergistic influence of doxorubicin and siRNA knockdown of the anti-apoptotic gene BCL-XL. Collectively, these studies highlight the broad utility of μ-tissue suspensions as an enabling approach for high n, populational analysis of 3D tissue biology in vitro and in vivo.
3-nitro-tyrosine as an internal quencher of autofluorescence enhances the compatibility of fluorescence based screening of OBOC combinatorial libraries.
Jared Townsend, Andrew Do, Alan Lehman, Seth Dixon, Babak Sanii and Kit S. Lam* June 01, 2010 Comb Chem High Throughput Screen. 2010 Jun;13(5):422-9.
View Abstract3-nitro-tyrosine as an internal quencher of autofluorescence enhances the compatibility of fluorescence based screening of OBOC combinatorial libraries.
Division of Hematology & Oncology, Department of Internal Medicine, UC Davis Cancer Center, University of California Davis, California, USA
Accurate MALDI-TOF/TOF sequencing of one-bead-one-compound peptide libraries with application to the identification of multiligand protein affinity agents using in situ click chemistry screening.
Lee SS, Lim J, Tan S, Cha J, Yeo SY, Agnew HD, Heath JR. January 15, 2010 Anal Chem 2010 Jan 15;82(2):672-9
View AbstractAccurate MALDI-TOF/TOF sequencing of one-bead-one-compound peptide libraries with application to the identification of multiligand protein affinity agents using in situ click chemistry screening.
Institute of Bioengineering and Nanotechnology, 31 Biopolis Way, The Nanos, Singapore 138669
Spectral Measurements of Large Particles by Flow Cytometry
Dakota A. Watson,¹ Daniel F. Gaskill,¹ Leif O. Brown,² Stephen K. Doorn,²and John P. Nolan¹* May 01, 2009 Cytometry A. 2009 May; 75(5): 460–464. doi: 10.1002/cyto.a.20706. 1) La Jolla Bioengineering Institute, La Jolla CA 92037 2) Los Alamos National Laboratory, Los Alamos NM 97545
View AbstractSpectral Measurements of Large Particles by Flow Cytometry
Flow cytometers designed to analyze large particles are enabling new applications in biology and chemistry. Likewise, flow spectroscopy approaches are extending the capabilities of the flow cytometry platform. Here we report on the adaptation of a commercial large particle analyzer to measure fluorescence and Raman spectra of individual particles at high speeds. We modified a Union Biometrica COPAS Plus instrument to allow red excitation and optical fiber-based light collection and spectral analysis using a spectrograph and CCD array detector. These modifications did not compromise the ability of the instrument to resolve different sized particles based on their extinction and time of flight signals. The modified instrument has the sensitivity and spectral resolution to measure the fluorescence and Raman signals from individual particles with signal integration times of 10 usec. The high speed spectral analysis of individual particles in flow will enable new applications in biological and chemical analyses.
Novel method for high-throughput colony PCR screening in nanoliter-reactors
Marcel Walser¹, Rene Pellaux¹, Andreas Meyer¹, Matthias Bechtold¹, Herve Vanderschuren², Richard Reinhardt³, Joseph Magyar4, Sven Panke¹ and Martin Held¹,* February 26, 2009 Nucl. Acids Res. (2009) 37 (8): e57/1-e57/8
View AbstractNovel method for high-throughput colony PCR screening in nanoliter-reactors
1) ETH Zurich, Institute of Process Engineering, BioProcess Laboratory (BPL), 2) ETH Zurich, Institute of Plant Science, Plant Biotechnology, Zurich, Switzerland, 3) Max Planck Institute for Molecular Genetics, Analytics & Computing, Berlin, Germany and 4) Harlan Laboratories Ltd., Environmental Safety and Metabolism, Itingen, Switzerland
Isolation of monoclonal microcarriers colonized by fluorescent E. coli
Walser M, Leibundgut RM, Pellaux R, Panke S, Held M. June 16, 2008 ETH Zurich, Institute of Process Engineering, BioProcess Laboratory, Universitätsstr. 6/CAB H88, CH-8092 Zurich, Switzerland.
View AbstractIsolation of monoclonal microcarriers colonized by fluorescent E. coli
Microencapsulation can be used for high-throughput (HT) processing of bacterial libraries. Ideally, each microcarrier would contain exactly one cell or colony. However, synthesis yields a mixture of capsules containing the desired one cell, multiple cells and empty microcarriers. E. coli cells expressing green fluorescent protein (GFP) were encapsulated in 35 nl Hydrogel carriers. HT sorting procedures by a COAPS sorter were then used to enrich samples to 95% monoclonality.
Published Online: Jun 16 2008 2:41PM; DOI: 10.1002/cyto.a.20597; Cytometry A. 2008 Sep;73(9):788-98.
Aminodeoxychorismate Synthase Inhibitors from One-Bead One-Compound Combinatorial Libraries: “Staged” Inhibitor Design
Seth Dixon,† Kristin T. Ziebart,† Ze He,‡ Mellisa Jeddeloh,† Choong Leol Yoo,† Xiaobing Wang,§ Alan Lehman,§ Kit S. Lam, § Michael D. Toney,*,† and Mark J. Kurth*,† November 10, 2006 J. Med. Chem., 49 (25), 7413 -7426, 2006. 10.1021/jm0609869 S0022-2623(06)00986-1
View AbstractAminodeoxychorismate Synthase Inhibitors from One-Bead One-Compound Combinatorial Libraries: “Staged” Inhibitor Design
†Department of Chemistry, University of California, One Shields Avenue, Davis, California 95616, ‡Department of Chemistry, Central Connecticut State University, 1615 Stanley Street, New Britain, Connecticut 06050, and §Department of Internal Medicine, Division of Hematology and Oncology, UC Davis Cancer Center, 4501 X Street, Sacramento, California 95817
SIRT1 Top 40 Hits: Use of One-Bead, One-Compound Acetyl-Peptide Libraries and Quantum Dots to Probe Deacetylase Specificity.
Adam L. Garske and John M. Denu December 13, 2005 Biochemistry 45(1) pp 94 – 101 (2006)
View AbstractSIRT1 Top 40 Hits: Use of One-Bead, One-Compound Acetyl-Peptide Libraries and Quantum Dots to Probe Deacetylase Specificity.
Departments of Biomolecular Chemistry and Chemistry, University of Wisconsin, Madison, Wisconsin 53706
OBOC Small-Molecule Combinatorial Library Encoded by Halogenated Mass-Tags
Sung Hee Hwang, Alan Lehman, Xin Cong, Marilyn M. Olmstead, Kit S. Lam, Carlito B. Lebrilla, and Mark J. Kurth September 17, 2004 Organic Letters, Volume 6(21), Pages 3829-3832 Department of Chemistry, University of California-Davis, One Shields Avenue, Davis, California 95616-5295, and Division of Hematology and Oncology, Department of Internal Medicine, University of California-Davis Cancer Center, 4501 X Street, Sacramento, California 95817
View AbstractOBOC Small-Molecule Combinatorial Library Encoded by Halogenated Mass-Tags
A bromine-/chlorine-containing mass-tag encoding strategy for a small-molecule OBOC combinatorial library is reported. The resulting MALDI FTMS isotope pattern of each tag clearly defines the component building blocks of each “hit” bead in an 1890-member demonstration library screened on-bead for binding against streptavidin via both enzyme-linked colorimetric and Quantum Dot/COPAS assays.
Combinatorial library of peptidotriazoles: identification of [1,2,3]-triazole inhibitors against a recombinant Leishmania mexicana cysteine protease.
Tornoe CW, Sanderson SJ, Mottram JC, Coombs GH, Meldal M. May 01, 2004 J Comb Chem. 2004 May-Jun;6(3):312-24.
View AbstractCombinatorial library of peptidotriazoles: identification of [1,2,3]-triazole inhibitors against a recombinant Leishmania mexicana cysteine protease.
Center for Solid-Phase Organic Combinatorial Chemistry, Department of Chemistry, Carlsberg Laboratory, Gamle Carlsberg Vej 10, DK-2500 Valby, Denmark.
Solid Phase Combinatorial Library of 1,3-Azole Containing Peptides for the Discovery of Matrix Metallo Proteinase Inhibitors
Caspar Christensen1, Christine Bruun Schiødt 2, Niels Tækker Foged2, Morten Meldal1 * December 08, 2003 QSAR & Combinatorial Science, Volume 22, Issue 7, Pages 754-766
View AbstractSolid Phase Combinatorial Library of 1,3-Azole Containing Peptides for the Discovery of Matrix Metallo Proteinase Inhibitors
1) Carlsberg Laboratory, Department of Chemistry, Gamle Carlsberg Vej 10, DK-2500 Valby, Denmark, 2) Nordic Bioscience A/S, Herlev Hovedgade 207, DK-2730 Herlev, Denmark.
New Technology Automates Sorting of Large, Bead-based CombiChem Libraries, P127.
18th American Peptide Symposium, July 19-23, 2003
K. Ver Donck1, L. Bols1, R. Bongaarts1, P. Van Osta1, A. J. Brouwer2,
R. M. J. Liskamp2, D. Perrault3, K. Kalutkiewicz3, J. Geysen1, and R. Pulak3
July 19, 2003
New Technology Automates Sorting of Large, Bead-based CombiChem Libraries, P127.
Union Biometrica-(Geel, BE1, Holliston, MA USA3), Utrecht University2 (Utrecht, NL).
From synthetic receptors towards synthetic antibodies using combinatorial chemistry, IN4.
2nd European Symposium on Combinatorial Sciences (Eurocombi 2), June 29-July 3
Rob M.J. Liskamp, Arwin J. Brouwer, Christina Chamorro, Menno C.F. Monnee, Till Opatz, Roland J. Pieters, Dirk T.S. Rijkers, Kirk-Jan van Zoelen.
June 29, 2003
From synthetic receptors towards synthetic antibodies using combinatorial chemistry, IN4.
Department of Medicinal Chemistry, Utrecht Institute for Pharmaceutical Sciences, Utrecht University, (Utrecht, The Netherlands).
Protein affinity profiling using solid-phase combinatorial libraries, O19.
2nd European Symposium on Combinatorial Sciences (Eurocombi 2), June 29-July 3
Anita J. Mathiesen, Sheryl Surve, Haifeng Yin, Phaedria M. St.Hilaire.
June 29, 2003
Protein affinity profiling using solid-phase combinatorial libraries, O19.
Carlsberg Biosectro, Valby (Denmark).
Bead-Based assays using COPAS™ Flow Sorting, P16.
2nd European Symposium on Combinatorial Sciences (Eurocombi 2), June 29-July 3
K. Ver Donck1, L. Bols1, R. Bongaarts1, P. Van Osta1, A. J. Brouwer2,
R. M. J. Liskamp2, D. Perrault3, J. Geysen1, and R. Pulak3
June 29, 2003
Bead-Based assays using COPAS™ Flow Sorting, P16.
Union Biometrica-(Geel, BE1, Holliston, MA USA3), Utrecht University2 (Utrecht, NL).
Bead Based Assays Using COPAS Flow Sorting, Practical Session
Innovative Combinatorial Approaches and Technologies, Practical Training Course (Florence, Italy) April 9-11, 2003
R. Bongaarts and A. Comas.
April 09, 2003
Bead Based Assays Using COPAS Flow Sorting, Practical Session
Union Biometrica, Geel, Belgium.
New technology automates sorting of large, bead--based combinatorial chemistry libraries, #87.
Lab Automation 2003, February 1-5, 2003, Palm Springs, CA
K. VerDonck1, L. Bols1, R. Bongaarts1, P. Van Osta1, J. Brouwer2, R. M. J. Liskamp2, J. Geysen1, D. Perrault3, K. Kalutkiewicz3, R. Pulak3
February 01, 2003
New technology automates sorting of large, bead--based combinatorial chemistry libraries, #87.
Union Biometrica 1, 3, (Geel, BE 1; Holliston, MA 3), Utrecht University (Utrecht, NL 2)
Assays for Bead-Based Technologies Using COPAS™ Flow Sorting, Session 3, Abstract 11.
ISLAR 2002, October 20-23, 2002
K. Ver Donck1, L. Bols1, R. Bongaarts1, P. Van Osta1, A. J. Brouwer2, R. M. J. Liskamp2, J. Geysen1, D. Perrault3, K. Kalutkiewicz3, R. Pulak3
October 20, 2002
Assays for Bead-Based Technologies Using COPAS™ Flow Sorting, Session 3, Abstract 11.
Union Biometrica-ESO 1, (Geel, BE), Union Biometrica 3, (Holliston, MA) Utrecht University 2 (Utrecht, NL)
The one-bead two-compound assay for solid-phase screening of Combinatorial libraries.
Morton Meldal September 16, 2002 Biopolymers, 66(2) 93-100.
View AbstractThe one-bead two-compound assay for solid-phase screening of Combinatorial libraries.
Carlsberg Laboratory, Department of Chemistry, Gamle Carlsberg Vej 10, DK-2500 Valby, Denmark.
At the higher end of High Content Screening: The MIAS HTS imaging device, #82.
MipTec-ICAR, May 27-30, 2002
Kris Ver Donck, Luc Bols, Peter Van Osta, Johan Geysen
May 27, 2002
At the higher end of High Content Screening: The MIAS HTS imaging device, #82.
Union Biometrica, Cipalstraat 3, B-2440 Geel, Belgium
Firms Creating More Focused Compound Libraries, Saving Time and Money in Drug Discovery Research
Nina Flanagan November 15, 2001 Genetic Engineering News, Vol. 21, No. 20, November 15, 2001
Firms Creating More Focused Compound Libraries, Saving Time and Money in Drug Discovery Research
Novel Methods for Analysis of Solid Phase Combinatorial Libraries on PEG-based Resins, #1134.
84th CSC Conference and Exhibition, May 2001
M. Meldal*, C. Gotfredsen, A. Papanikos, A. Meissner, M. Grotli, T. Groth, J. Beyer, J. Duus, C. Tornoe, and P. St. Hilaire
May 01, 2001
Novel Methods for Analysis of Solid Phase Combinatorial Libraries on PEG-based Resins, #1134.
SPOCC-center, Carlsberg Laboratory, Department of Chemistry, Gamle Carlsberg Vej 10, DK-2500 Valby, Denmark.