Development and Application of Activity-based Fluorescent Probes for High-Throughput Screening

2021 ◽  
Vol 28 ◽  
Author(s):  
Juan Cheng ◽  
Xin Li
Keyword(s):  
Drug Development ◽  
High Throughput ◽  
Fluorescent Probes ◽  
Analytical Methods ◽  
High Throughput Screening ◽  
High Sensitivity ◽  
Rapid Identification ◽  
Screening Assays ◽  
Hit Identification ◽  
Working Mechanisms

: High-throughput screening facilitates the rapid identification of novel hit compounds; however, it remains challenging to design effective high-throughput assays, partially due to the difficulty of achieving sensitivity in the assay techniques. Among the various analytical methods that are used, fluorescence-based assays dominate owing to their high sensitivity and ease of operation. Recent advances in activity-based sensing/imaging have further expanded the availability of fluorescent probes as monitors for high-throughput screening of result outputs. In this study, we have reviewed various activity-based fluorescent probes used in high-throughput screening assays, emphasizing their structure-related working mechanisms. Moreover, we have explored the possibility of the development of additional and better probes to boost hit identification and drug development against various targets.

Application of AlphaLISA in bioanalysis

2020 ◽  
pp. 16-37
Author(s):  
Stanislav Cherepushkin
Keyword(s):  
Drug Development ◽  
Ligand Binding ◽  
High Throughput ◽  
Protein Interactions ◽  
High Throughput Screening ◽  
High Sensitivity ◽  
Medical Diagnostics ◽  
Protein Protein Interactions ◽  
Binding Assays ◽  
Post Translational Modifications

The use and development of biotherapeutics increases and the need for accurate, sensitive and robust bioanalytical methods is also increasing. ELISA and other ligand-binding assays are the most widely used methods for the quantification of macromolecules in complex biological samples. One of the alternatives to ELISA is AlphaLISA — a versatile chemiluminescent ligand binding assay using a homogeneous no-wash protocol. AlphaLISA assays are suited for automation and exhibit high sensitivity, high throughput and wide analytical range. Since the early 2000s, this method has been used in science, medicine, and drug development for wide variety of applications, including the quantification of analytes, immunogenicity, protein-protein interactions, enzyme activity, post-translational modifications and epigenetics. In this review, we describe the principles of the AlphaLISA assay and its application in bioanalytical studies (pharmacokinetics and immunogenicity) and high-throughput screening in drug development, medical diagnostics and pathogens detection.

A Simple Statistical Parameter for Use in Evaluation and Validation of High Throughput Screening Assays

1999 ◽  
Vol 4 (2) ◽  
pp. 67-73 ◽  
Author(s):  
Ji-Hu Zhang ◽  
Thomas D. Y. Chung ◽  
Kevin R. Oldenburg
Keyword(s):  
High Throughput ◽  
High Throughput Screening ◽  
Dynamic Range ◽  
Statistical Parameter ◽  
Assay Optimization ◽  
Screening Assays ◽  
Signal Dynamic Range ◽  
Hit Identification ◽  
Data Variation

The ability to identify active compounds ("hits") from large chemical libraries accurately and rapidly has been the ultimate goal in developing high-throughput screening (HTS) assays. The ability to identify hits from a particular HTS assay depends largely on the suitability or quality of the assay used in the screening. The criteria or parameters for evaluating the "suitability" of an HTS assay for hit identification are not well defined and hence it still remains difficult to compare the quality of assays directly. In this report, a screening window coefficient, called "Z- factor," is defined. This coefficient is reflective of both the assay signal dynamic range and the data variation associated with the signal measurements, and therefore is suitable for assay quality assessment. The Z-factor is a dimensionless, simple statistical characteristic for each HTS assay. The Z-factor provides a useful tool for comparison and evaluation of the quality of assays, and can be utilized in assay optimization and validation.

scholarly journals Action plan for hit identification (APHID): KAT6A as a case study

2020 ◽  
Vol 12 (5) ◽  
pp. 423-437 ◽  
Author(s):  
Xiangyan Yi ◽  
Lian Xue ◽  
Tim Thomas ◽  
Jonathan B Baell
Keyword(s):  
High Throughput ◽  
High Throughput Screening ◽  
Action Plan ◽  
Hit Identification

Here, we describe our action plan for hit identification (APHID) that guides the process of hit triage, with elimination of less tractable hits and retention of more tractable hits. We exemplify the process with reference to our high-throughput screening (HTS) campaign against the enzyme, KAT6A, that resulted in successful identification of a tractable hit. We hope that APHID could serve as a useful, concise and digestible guide for those involved in HTS and hit triage, especially those that are relatively new to this exciting and continually evolving technology.

scholarly journals Design of High-Throughput Screening Assays and Identification of a SUMO1-Specific Small Molecule Chemotype Targeting the SUMO-Interacting Motif-Binding Surface

2015 ◽  
Vol 17 (4) ◽  
pp. 239-246 ◽  
Author(s):  
Aileen Y. Alontaga ◽  
Yifei Li ◽  
Chih-Hong Chen ◽  
Chen-Ting Ma ◽  
Siobhan Malany ◽  
...  
Keyword(s):  
Small Molecule ◽  
High Throughput ◽  
High Throughput Screening ◽  
Binding Surface ◽  
Screening Assays

scholarly journals High-Throughput Screening and Drug Development to Fix RyR Pathology

2021 ◽  
Vol 120 (3) ◽  
pp. 148a-149a
Author(s):  
Robyn T. Rebbeck ◽  
Kaja Rozman ◽  
Gabrielle M. Evans ◽  
Jacob Schwarz ◽  
Marzena Baran ◽  
...  
Keyword(s):  
Drug Development ◽  
High Throughput ◽  
High Throughput Screening
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