Nucleic Acids Research, Vol 24, Issue 4 702-706, Copyright © 1996 by Oxford University Press
KA Davis, B Abrams, Y Lin and SD Jayasena
To investigate the feasibility of using oligonucleotides in flow cytometry
we describe a model system consisting of human neutrophil elastase (HNE)
coated on 3.3 micro beads and a high affinity DNA ligand for HNE isolated
by in vitro selection (SELEX). In this system the fluoresceinated DNA
ligand was equally effective as an anti- HNE antibody in detecting HNE on
beads. The location on and the chemistry of attachment of fluorescein to
the DNA ligand is critical for the sensitivity of detection. DNA constructs
in which fluorescein was conjugated via an ethylene glycol tether to either
the 5'-end or near the 3'-end gave much higher signals than did probes with
fluorescein directly conjugated to either end. Second-step staining with
strepavidin-conjugated phycoerythrin was accomplished using a biotinylated
DNA ligand in the initial staining of HNE beads. These data suggest that
instead of, or in addition to, antibodies high affinity oligonucleotide
probes can be useful in diagnostic applications based on flow cytometry.
ARTICLES
Use of a high affinity DNA ligand in flow cytometry
Becton Dickinson Immunocytometry Systems, San Jose, CA 95131 USA.
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