Nucleic Acids Research, Vol 26, Issue 17 3915-3924, Copyright © 1998 by Oxford University Press
KA Davis, Y Lin, B Abrams and SD Jayasena
We have used recombinant human CD4 presented on beads as an affinity matrix
to screen a 2'-F-pyrimidine-containing RNA library with a complexity of
approximately 10(14) molecules. Affinity-selected aptamers bind recombinant
CD4 with low nanomolar equilibrium dissociation constants. These
high-affinity aptamers conjugated to different fluorophores such as
fluorescein and phycoerythrin were used to stain cells, expressing human
CD4 on cell surface, for analysis by flow cytometry. Aptamers, conjugated
to fluorophores, stained mouse T cells that express human CD4 on the
surface, but not the control mouse T cells lacking human CD4. The control
cells, however, do express mouse CD4 whose extracellular domain has 55%
sequence identity to the human form. These human CD4-specific aptamers
selectively stained CD4(+) T cells in a preparation of human peripheral
blood mononuclear cells. These results and others suggest that aptamers are
emerging as a versatile class of molecules that can be used for various
diagnostic applications performed under different formats or platforms.
ARTICLES
Staining of cell surface human CD4 with 2'-F-pyrimidine-containing RNA aptamers for flow cytometry
NeXstar Pharmaceuticals Inc., 2860 Wilderness Place, Boulder, CO 80301, USA and Becton Dickinson Immunocytometry Systems, 2350 Qume Drive, San Jose, CA 95131, USA.
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