دورية أكاديمية
Bioconjugation of Antibodies and Enzyme Labels onto Magnetic Beads.
| العنوان: | Bioconjugation of Antibodies and Enzyme Labels onto Magnetic Beads. |
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| المؤلفون: | Otieno BA; Department of Chemistry, University of Connecticut, Storrs, Connecticut, USA., Krause CE; Department of Chemistry, University of Hartford, West Hartford, Connecticut, USA., Rusling JF; Department of Chemistry, University of Connecticut, Storrs, Connecticut, USA; Institute of Materials Science, University of Connecticut, Storrs, Connecticut, USA; Department of Cell Biology, University of Connecticut Health Center, Farmington, Connecticut, USA; School of Chemistry, National University of Ireland, Galway, Ireland. Electronic address: james.rusling@uconn.edu. |
| المصدر: | Methods in enzymology [Methods Enzymol] 2016; Vol. 571, pp. 135-50. Date of Electronic Publication: 2016 Jan 19. |
| نوع المنشور: | Journal Article; Research Support, N.I.H., Extramural |
| اللغة: | English |
| بيانات الدورية: | Publisher: Academic Press Country of Publication: United States NLM ID: 0212271 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1557-7988 (Electronic) Linking ISSN: 00766879 NLM ISO Abbreviation: Methods Enzymol Subsets: MEDLINE |
| أسماء مطبوعة: | Original Publication: New York, Academic Press. |
| مواضيع طبية MeSH: | Biosensing Techniques*, Antibodies, Immobilized/*chemistry , Enzymes, Immobilized/*chemistry, Affinity Labels/chemistry ; Horseradish Peroxidase/chemistry ; Immunoassay ; Magnets/chemistry ; Microfluidic Analytical Techniques ; Nanoparticles/chemistry ; Streptavidin/chemistry |
| مستخلص: | Immunoassays employ antibodies and labels to capture and detect target macromolecular analytes, often from complex sample matrices such as serum, plasma, or saliva. The high affinity and specificity of antibody-antigen interactions makes immunoassays critically important analytical techniques for clinical diagnostics as well as other research applications in the areas of pharmaceutical and environmental analysis. Integration of magnetic beads (MBs) into immunoassays and other bioanalytical methodologies is a valuable approach to allow efficient target capture, enrichment, and convenient separation. In addition, large signal amplification can be achieved by preconcentration of the target and by attaching many thousands of enzyme labels to the MBs. These features have enabled MB-based biosensors to achieve ultra-low detection limits needed for advanced clinical diagnostics that are challenging or impossible using traditional immunoassays. MBs are employed either as mobile substrates for target analyte capture, as detection labels (or label carriers), or simultaneously as substrates and labels. For optimal assay performance, it is crucial to apply an easy, efficient, and robust bead-probe conjugation protocol, and to thoroughly characterize the bioconjugated products. Herein, we describe methods used in our laboratory to functionalize MBs with antibodies and enzyme labels for ultrasensitive detection of protein analytes. We also present detailed strategies for characterizing the MB bioconjugates. (© 2016 Elsevier Inc. All rights reserved.) |
| معلومات مُعتمدة: | R01 EB016707 United States EB NIBIB NIH HHS; R01 EB014586 United States EB NIBIB NIH HHS |
| فهرسة مساهمة: | Keywords: Antibodies; Bicinchoninic acid assay; Bioconjugation; Enzyme kinetics; Enzymes; Immunoassay; Magnetic beads |
| المشرفين على المادة: | 0 (Affinity Labels) 0 (Antibodies, Immobilized) 0 (Enzymes, Immobilized) 9013-20-1 (Streptavidin) EC 1.11.1.- (Horseradish Peroxidase) |
| تواريخ الأحداث: | Date Created: 20160427 Date Completed: 20170807 Latest Revision: 20220408 |
| رمز التحديث: | 20221213 |
| DOI: | 10.1016/bs.mie.2015.10.005 |
| PMID: | 27112398 |
| قاعدة البيانات: | MEDLINE |
Full Text Finder | تدمد: | 1557-7988 |
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| DOI: | 10.1016/bs.mie.2015.10.005 |