Abstract
Receptor-mediated cellular uptake of physiological regulators such as nutritional and hormonal factors represents a transport event with important consequences for cell differentiation, death, or proliferation. Although internalization pathways are important points of regulation, they have not been extensively explored as pharmacological targets in most cell types. An experimental strategy based on ligand-enzyme conjugates is presented in this report that may facilitate high-throughput screening for potent chemical modulators of the transport events. The method was tested on a human epidermoid cell line using a streptavidin-peroxidase conjugate, and human holo-transferrin as the model ligand, in a biotin-ligand conjugate. The proposed screening method is rapid, can be performed using multi-well plates, and involves small assay volumes. The modular nature of the ligand complex makes this method adaptable to the use of other biotinylated ligands, and the use of avidins conjugated to other enzymes. As is discussed, the method may also be applicable to other in vitro and in vivo transport assays.
Keywords: Cell transport, epidermoid cells, ligand conjugates, receptor-mediated endocytosis, screening assay, Endocytosis, clathrin-mediated endocytosis, transferrin endocytosis, transferrin RME model, ligand-enzyme conjugate assay.
Combinatorial Chemistry & High Throughput Screening
Title:Ligand-Peroxidase Conjugates for Quantification of Receptor-Mediated Transport into Cells
Volume: 16 Issue: 4
Author(s): Jie Chen and Amandio Vieira
Affiliation:
Keywords: Cell transport, epidermoid cells, ligand conjugates, receptor-mediated endocytosis, screening assay, Endocytosis, clathrin-mediated endocytosis, transferrin endocytosis, transferrin RME model, ligand-enzyme conjugate assay.
Abstract: Receptor-mediated cellular uptake of physiological regulators such as nutritional and hormonal factors represents a transport event with important consequences for cell differentiation, death, or proliferation. Although internalization pathways are important points of regulation, they have not been extensively explored as pharmacological targets in most cell types. An experimental strategy based on ligand-enzyme conjugates is presented in this report that may facilitate high-throughput screening for potent chemical modulators of the transport events. The method was tested on a human epidermoid cell line using a streptavidin-peroxidase conjugate, and human holo-transferrin as the model ligand, in a biotin-ligand conjugate. The proposed screening method is rapid, can be performed using multi-well plates, and involves small assay volumes. The modular nature of the ligand complex makes this method adaptable to the use of other biotinylated ligands, and the use of avidins conjugated to other enzymes. As is discussed, the method may also be applicable to other in vitro and in vivo transport assays.
Export Options
About this article
Cite this article as:
Chen Jie and Vieira Amandio, Ligand-Peroxidase Conjugates for Quantification of Receptor-Mediated Transport into Cells, Combinatorial Chemistry & High Throughput Screening 2013; 16 (4) . https://dx.doi.org/10.2174/1386207311316040005
DOI https://dx.doi.org/10.2174/1386207311316040005 |
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
- 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
-
Breast Cancer: Current Developments in Molecular Approaches to Diagnosis and Treatment
Recent Patents on Anti-Cancer Drug Discovery Tubulin-Targeting Agents in Hybrid Drugs
Current Medicinal Chemistry Combination Treatment of Glioblastoma by Low-Dose Radiation and Genistein
Current Radiopharmaceuticals The Importance of TLR3 Expression and Hormonal Regulation of TLR3- Induced Immune Responses in the Human Endometrium
Current Immunology Reviews (Discontinued) A Review on the Role of Phytosterols: New Insights Into Cardiovascular Risk
Current Pharmaceutical Design miR-15b and miR-21 as Circulating Biomarkers for Diagnosis of Glioma
Current Genomics New Insights about the Potential Application of the Association of Vitamins C (Sodium Ascorbate) and K3 (Menadione) as Auxiliary Therapy in Cancer Treatment
Medicinal Chemistry Reviews - Online (Discontinued) Biosystems Engineering of Prokaryotes with Tumor-Killing Capacities
Current Pharmaceutical Design Mechanisms of CTLA-4-Ig in Tolerance Induction
Current Pharmaceutical Design Immunoproteasome-Selective Inhibitors: A Promising Strategy to Treat Hematologic Malignancies, Autoimmune and Inflammatory Diseases
Current Medicinal Chemistry Synergistic Effect of α-Solanine and Cisplatin Induces Apoptosis and Enhances Cell Cycle Arrest in Human Hepatocellular Carcinoma Cells
Anti-Cancer Agents in Medicinal Chemistry Revisiting the ABCs of Multidrug Resistance in Cancer Chemotherapy
Current Pharmaceutical Biotechnology Pharmacological Interventions for the Prevention and Treatment of Kidney Injury Induced by Radiotherapy: Molecular Mechanisms and Clinical Perspectives
Current Molecular Pharmacology Nucleic Acid Aptamers Against Protein Kinases
Current Medicinal Chemistry In-Situ Hybridization as a Molecular Tool in Cancer Diagnosis and Treatment
Current Medicinal Chemistry Genetic Abnormalities in Prostate Cancer
Current Genomics Extracellular Vesicles as Innovative Tools for Assessing Adverse Effects of Immunosuppressant Drugs
Current Medicinal Chemistry Preclinical Evaluation of New Taxoids
Current Pharmaceutical Design Proteomic Analysis of Neutrophil Priming by PAF
Protein & Peptide Letters Specific Targeting of Akt Kinase Isoforms: Taking the Precise Path for Prevention and Treatment of Cancer
Current Drug Targets