Abstract
In the one-bead-one-compound (OBOC) combinatorial method, compounds are constructed on bead resin via split-mix library synthesis such that multiple copies of the same compound are displayed on each bead. These libraries are rapidly screened with enzyme-linked colorimetric, fluorescent, radiometric, or whole-cell binding assays. While fluorescence-based probes are powerful tools in OBOC screening, their utility is greatly limited by the intrinsic fluorescence of many commonly used solid supports, (e.g., TentaGel), residual coupling reagents, and library compounds. To overcome this problem, we topologically partitioned TentaGel resin with a thin Fmoc-protected outer layer and an unprotected inner core. The inner core was derivatized with 3-nitro-tyrosine, followed by random peptide library construction. Spectral scans from a confocal microscope showed a dramatic decrease in the autofluorescence of blank beads and OBOC peptide libraries across a broad range of the optical spectrum. The quenching capacity of 3-nitro-tyrosine was also visualized in fluorescent micrographs. Using biotin/streptavidin as a model ligand/receptor system, we demonstrated a marked increase in visibility of three commercially available fluorescent probes binding to quenched beads, and increased feasibility of using a robust and efficient fluorescence-based, bead sorting platform known as COPAS™. These data show that using 3-nitro-tyrosine as an internal quencher greatly enhances the compatibility of fluorescence-based applications and OBOC combinatorial screening.
Keywords: OBOC library, combinatorial chemistry, high throughput screening, fluorescence-based screening, fluorescent quenching, 3-nitro-tyrosine, autofluorescence, quantum dot
Combinatorial Chemistry & High Throughput Screening
Title: 3-Nitro-Tyrosine as an Internal Quencher of Autofluorescence Enhances the Compatibility of Fluorescence Based Screening of OBOC Combinatorial Libraries
Volume: 13 Issue: 5
Author(s): Jared Townsend, Andrew Do, Alan Lehman, Seth Dixon, Babak Sanii and Kit S. Lam
Affiliation:
Keywords: OBOC library, combinatorial chemistry, high throughput screening, fluorescence-based screening, fluorescent quenching, 3-nitro-tyrosine, autofluorescence, quantum dot
Abstract: In the one-bead-one-compound (OBOC) combinatorial method, compounds are constructed on bead resin via split-mix library synthesis such that multiple copies of the same compound are displayed on each bead. These libraries are rapidly screened with enzyme-linked colorimetric, fluorescent, radiometric, or whole-cell binding assays. While fluorescence-based probes are powerful tools in OBOC screening, their utility is greatly limited by the intrinsic fluorescence of many commonly used solid supports, (e.g., TentaGel), residual coupling reagents, and library compounds. To overcome this problem, we topologically partitioned TentaGel resin with a thin Fmoc-protected outer layer and an unprotected inner core. The inner core was derivatized with 3-nitro-tyrosine, followed by random peptide library construction. Spectral scans from a confocal microscope showed a dramatic decrease in the autofluorescence of blank beads and OBOC peptide libraries across a broad range of the optical spectrum. The quenching capacity of 3-nitro-tyrosine was also visualized in fluorescent micrographs. Using biotin/streptavidin as a model ligand/receptor system, we demonstrated a marked increase in visibility of three commercially available fluorescent probes binding to quenched beads, and increased feasibility of using a robust and efficient fluorescence-based, bead sorting platform known as COPAS™. These data show that using 3-nitro-tyrosine as an internal quencher greatly enhances the compatibility of fluorescence-based applications and OBOC combinatorial screening.
Export Options
About this article
Cite this article as:
Townsend Jared, Do Andrew, Lehman Alan, Dixon Seth, Sanii Babak and S. Lam Kit, 3-Nitro-Tyrosine as an Internal Quencher of Autofluorescence Enhances the Compatibility of Fluorescence Based Screening of OBOC Combinatorial Libraries, Combinatorial Chemistry & High Throughput Screening 2010; 13 (5) . https://dx.doi.org/10.2174/138620710791292994
DOI https://dx.doi.org/10.2174/138620710791292994 |
Print ISSN 1386-2073 |
Publisher Name Bentham Science Publisher |
Online ISSN 1875-5402 |
Call for Papers in Thematic Issues
Artificial Intelligence Methods for Biomedical, Biochemical and Bioinformatics Problems
Recently, a large number of technologies based on artificial intelligence have been developed and applied to solve a diverse range of problems in the areas of biomedical, biochemical and bioinformatics problems. By utilizing powerful computing resources and massive amounts of data, methods based on artificial intelligence can significantly improve the ...read more
Eco-friendly Agents for Biological Control of Pathogenic Diseases
The discovery of an alternative biological approach to disease management includes work on medicinal products derived from natural sources as a starting point for the development of eco-friendly agents for these diseases and the injuries they cause, as well as reducing human contact with hazardous chemicals and their residues. We ...read more
Emerging trends in diseases mechanisms, noble drug targets and therapeutic strategies: focus on immunological and inflammatory disorders
Recently infectious and inflammatory diseases have been a key concern worldwide due to tremendous morbidity and mortality world Wide. Recent, nCOVID-9 pandemic is a good example for the emerging infectious disease outbreak. The world is facing many emerging and re-emerging diseases out breaks at present however, there is huge lack ...read more
Exploring Spectral Graph Theory in Combinatorial Chemistry
Scope of the Thematic Issue: Combinatorial chemistry involves the synthesis and analysis of a large number of diverse compounds simultaneously. Traditional methods rely on brute force experimentation, which can be time-consuming and resource-intensive. Spectral Graph Theory, a branch of mathematics dealing with the properties of graphs in relation to the ...read more
![](/images/wayfinder.jpg)
- Author Guidelines
- Graphical Abstracts
- Fabricating and Stating False Information
- Research Misconduct
- Post Publication Discussions and Corrections
- Publishing Ethics and Rectitude
- Increase Visibility of Your Article
- Archiving Policies
- Peer Review Workflow
- Order Your Article Before Print
- Promote Your Article
- Manuscript Transfer Facility
- Editorial Policies
- Allegations from Whistleblowers
Related Articles
-
Persistent Current Blockers of Voltage-Gated Sodium Channels: A Clinical Opportunity for Controlling Metastatic Disease
Recent Patents on Anti-Cancer Drug Discovery Impact of GSTT1 and GSTM1 Polymorphisms in the Susceptibility to Philadelphia Negative Chronic Myeloid Leukaemia
Current Cancer Drug Targets Molecular Phenotype of CXCL12β 3UTR G801A Polymorphism (rs1801157) Associated to HIV-1 Disease Progression
Current HIV Research Baseline CD4/CD8 T-Cell Ratio Predicts Prompt Immune Restoration Upon cART Initiation
Current HIV Research Expansion of Umbilical Cord Blood for Clinical Transplantation
Current Stem Cell Research & Therapy Cancer Estimates in Brazil Reveal Progress for the Most Lethal Malignancies
Current Topics in Medicinal Chemistry p53-Mdm2 Interaction Inhibitors as Novel Nongenotoxic Anticancer Agents
Current Cancer Drug Targets Current status of Immunotherapy in B Cell Malignancies
Current Drug Targets Molecular Probing and Imaging of Histone Deacetylase Inhibitors in Cancer Treatment
Anti-Cancer Agents in Medicinal Chemistry The Health-promoting Potential of Edible Mushroom Proteins
Current Pharmaceutical Design Utilising Nanotechnology and Nanosystems for Treatment of Rare Diseases
Pharmaceutical Nanotechnology Marine-derived Natural Products as Anticancer Agents
Medicinal Chemistry Prevalence and Risk Factors of Vitamin D Deficiency in Critically Ill Patients
Inflammation & Allergy - Drug Targets (Discontinued) Monoclonal Antibodies in the Treatment of Systemic Lupus Erythematosus
Current Drug Targets Recent Design and Structure-Activity Relationship Studies on the Modifications of DHFR Inhibitors as Anticancer Agents
Current Medicinal Chemistry Application of Olefin Cross-Metathesis to the Synthesis of Biologically Active Natural Products
Current Topics in Medicinal Chemistry The link between Hepatic Vitamin A Metabolism and Nonalcoholic Fatty Liver Disease
Current Drug Targets Ubiquitin Carboxyl Hydrolase L1 Significance for Human Diseases
Protein & Peptide Letters The Role of Notch Signaling Pathway in Epithelial-Mesenchymal Transition (EMT) During Development and Tumor Aggressiveness
Current Drug Targets Anti-Inflammatory Drugs in Psychiatry
Inflammation & Allergy - Drug Targets (Discontinued)