High-throughput screening strategies for targeted identification of therapeutic compounds in colorectal cancer

Customized compound picking and plating of very large corporate screening decks (many 100,000s) for high-throughput screening is generally restricted, both from a time and cost perspective. Here we present a stratified screening deck with accompanying plating design for use with very large corporate compound collections. The deck is plated as a whole, but copies for screening can be downsized flexibly and quickly on the fly, without the need for repicking of physical samples. We show that such downsized sets maximize returns and yield results superior to randomly picked subsets of the same size. For the proposed stratified plating design, structurally diverse subsets that cover the full collection in terms of compound diversity and favorable compound properties can be produced economically and quickly from the full set of master plates. The design was implemented globally at AstraZeneca in 2009 and has enabled substantial cost-saving in screening campaigns, as set size requirements can be met on a per-screen basis, using a single, preplated master deck.

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Process Validation and Screen Reproducibility in High-Throughput Screening

CrossRef Listing of Deleted DOIs ◽

10.1177/1087057108326664 ◽

2008 ◽

Vol 14 (1) ◽

pp. 66-76 ◽

Cited By ~ 30

Author(s):

Isabel Coma ◽

Liz Clark ◽

Emilio Diez ◽

Gavin Harper ◽

Jesus Herranz ◽

...

Keyword(s):

Drug Discovery ◽

High Throughput ◽

High Throughput Screening ◽

Large Scale ◽

Academic Community ◽

Major Step ◽

Chemical Genomic ◽

Screening Strategies ◽

Medical Diagnostic ◽

Compound Screening

The use of large-scale compound screening has become a key component of drug discovery projects in both the pharmaceutical and the biotechnological industries. More recently, these activities have also been embraced by the academic community as a major tool for chemical genomic activities. High-throughput screening (HTS) activities constitute a major step in the initial drug discovery efforts and involve the use of large quantities of biological reagents, hundreds of thousands to millions of compounds, and the utilization of expensive equipment. All these factors make it very important to evaluate in advance of the HTS campaign any potential issues related to reproducibility of the experimentation and the quality of the results obtained at the end of these very costly activities. In this article, the authors describe how GlaxoSmithKline (GSK) has addressed the need of a true validation of the HTS process before embarking in full HTS campaigns. They present 2 different aspects of the so-called validation process: (1) optimization of the HTS workflow and its validation as a quality process and (2) the statistical evaluation of the HTS, focusing on the reproducibility of results and the ability to distinguish active from nonactive compounds in a vast collection of samples. The authors describe a variety of reproducibility indexes that are either innovative or have been adapted from generic medical diagnostic screening strategies. In addition, they exemplify how these validation tools have been implemented in a number of case studies at GSK. ( Journal of Biomolecular Screening 2009:66-76)

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High-Throughput Screening Strategies for Cardiac Ion Channels

Trends in Cardiovascular Medicine ◽

10.1016/s1050-1738(01)00086-x ◽

2001 ◽

Vol 11 (2) ◽

pp. 54-59 ◽

Cited By ~ 18

Author(s):

R Numann

Keyword(s):

Ion Channels ◽

High Throughput ◽

High Throughput Screening ◽

Cardiac Ion Channels ◽

Screening Strategies

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Microsatellite Instability in Colorectal Cancer: Considerations for Molecular Diagnosis and High-Throughput Screening of Archival Tissues

Clinical Chemistry ◽

10.1373/clinchem.2003.030700 ◽

2004 ◽

Vol 50 (6) ◽

pp. 1082-1086 ◽

Cited By ~ 15

Author(s):

Manuel Salto-Tellez ◽

Soo Chin Lee ◽

Lily L Chiu ◽

Chi Kuen Lee ◽

May Chin Yong ◽

...

Keyword(s):

Colorectal Cancer ◽

Microsatellite Instability ◽

Molecular Diagnosis ◽

High Throughput ◽

High Throughput Screening

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Mapping Out Receptors: High Throughput Screening of Endothelial Monocyte Activating Polypeptide II (EMAPII) Using PRESTO-TANGO

Proceedings of IMPRS ◽

10.18060/22721 ◽

2018 ◽

Vol 1 (1) ◽

Author(s):

Nathan Lam ◽

Bernhard Maier ◽

Sarvesh Chelvanambi ◽

Takashi Hato ◽

Matthias Clauss

Keyword(s):

Colorectal Cancer ◽

Cardiovascular Disease ◽

Receptor Binding ◽

High Throughput ◽

High Throughput Screening ◽

Neutralizing Antibodies ◽

Signal To Noise Ratio ◽

Receptor System ◽

Gpcr Activation ◽

Cxcr3 Ligand

Background: Secreted endothelial monocyte activating polypeptide II (EMAPII/AIMP1) is a pro-apoptotic, pro-inflammatory ligand implicated in diseases such as colorectal cancer, cardiovascular disease, and emphysema. Thus, EMAPII has been shown to induce apoptosis through CXCR3 receptor binding. However, not all EMAPII functions have been attributed to CXCR3. Discovery of new receptors interacting with EMAPII could lead to development of new therapies blocking cognate ligand-receptor binding. We hypothesize the existence of at least one unknown secondary receptor for EMAPII. Methods: The PRESTO-TANGO assay, a construct which converts G-protein coupled receptor (GPCR) ligand binding into luciferase activity measurable by luminometer, was validated using transfection with TANGO-modified CXCR3 and S1P1R as test receptors in HTLA cells. Protocols for cell transfection, adherence, and cultivation were optimized with IP10, EMAPII, and S1P as test ligands. Results: The assay was successfully validated using several GPCR activation readouts. Binding curves were generated for S1P/S1P1 (EC50= 569nM), IP10/CXCR3 (EC50= 47.1 nM), and EMAPII/CXCR3 (EC50= 628 nM). Conditions for the PRESTO-TANGO assay were further refined for maximal signal-to-noise ratio and robust inter-assay reproducibility in preparation for high-throughput screening. We are currently testing 314 TANGO-modified GPCRs for EMAPII affinity. Conclusion: We have validated the Tango assay for the known EMAPII-CXCR3 ligand-receptor system, a valuable tool for evaluation of anti-EMAPII therapeutics. Discovery of a novel EMAPII receptor would allow for the development of therapies including neutralizing antibodies (analogous to the PD1 receptor antibody Pembrolizumab for solid tumors) in diseases such as colorectal cancer, cardiovascular disease, and emphysema.

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