Your email was sent successfully. Check your inbox.

An error occurred while sending the email. Please try again.

Proceed reservation?

Export
  • 1
    Keywords: CELLS ; BLOOD ; CELL ; evaluation ; Germany ; SYSTEM ; SYSTEMS ; GENOME ; microarray ; PROTEIN ; PROTEINS ; TIME ; COMPLEX ; COMPLEXES ; T cell ; T cells ; T-CELL ; T-CELLS ; BIOLOGY ; MOLECULAR-BIOLOGY ; LINKAGE ; SIGNAL ; cytokines ; antibodies ; antibody ; PERFORMANCE ; AMPLIFICATION ; MICROARRAY DATA ; microarrays ; DESIGN ; PLASMA ; CANCER-CELLS ; PARAMETERS ; Jun ; STRATEGIES ; sensitivity ; MICROARRAY ANALYSIS ; INTERFERON-GAMMA ; IMMUNOASSAYS ; PROTEOMICS ; CYTOKINE ; molecular biology ; molecular ; CHEMISTRY ; ELISA ; monitoring ; RHEUMATOID-ARTHRITIS ; antibody microarray ; CYTOKINE PRODUCTION ; LEVEL ; analysis ; methods ; ROLLING-CIRCLE AMPLIFICATION ; EXTENT ; SPECIMENS ; MATTER ; protein profiling ; microspot kinetics ; MASS-TRANSPORT ; quantitative ; detection strategies ; INTERLEUKIN-4 PRODUCTION ; PLASMA-PROTEOME ; TRITON X-100
    Abstract: Antibody microarrays have often had limited success in detection of low abundant proteins in complex specimens. Signal amplification systems improve this situation, but still are quite laborious and expensive. However, the issue of sensitivity is more likely a matter of kinetically appropriate microarray design as demonstrated previously. Hence, we re-examined in this study the suitability of simple and inexpensive detection approaches for highly sensitive antibody microarray analysis. N-hydroxysuccinimidyl ester (NHS)- and Universal Linkage System (ULS)-based fluorescein and biotin labels used as tags for subsequent detection with anti-fluorescein and extravidin, respectively, as well as fluorescent dyes were applied for analysis of blood plasma. Parameters modifying strongly the performance of microarray detection such as labeling conditions, incubation time, concentrations of anti-fluorescein and extravidin and extent of protein labeling were analyzed and optimized in this study. Indirect detection strategies whether based on NHS- or ULS-chemistries strongly outperformed direct fluorescent labeling and enabled detection of low abundant cytokines with many dozen-fold signal-to-noise ratios. Finally, particularly sensitive detection chemistry was applied to monitoring cytokine production of stimulated peripheral T cells. Microarray data were in accord with quantitative cytokine levels measured by ELISA and Luminex, demonstrating comparable reliability and femtomolar range sensitivity of the established microarray approach
    Type of Publication: Journal article published
    PubMed ID: 17474144
    Signatur Availability
    BibTip Others were also interested in ...
Close ⊗
This website uses cookies and the analysis tool Matomo. More information can be found here...