PMID- 15801725
OWN - NLM
STAT- MEDLINE
DCOM- 20070213
LR  - 20151119
IS  - 0003-2700 (Print)
IS  - 0003-2700 (Linking)
VI  - 77
IP  - 7
DP  - 2005 Apr 1
TI  - DNA aptamer-based bioanalysis of IgE by fluorescence anisotropy.
PG  - 1963-70
AB  - A rapid, homogeneous aptamer-based bioanalysis is reported for the sensitive 
      detection of immunoglobulin E (IgE) using fluorescence polarization (FP). 
      5'-End-labeled D17.4 DNA aptamer was used for IgE detection based on the 
      anisotropy differences of the labeled ligand. Two different fluorophores, 
      fluorescein and Texas Red, were used to analyze IgE in the low-nanomolar range 
      with high specificity. Measurable anisotropy changes were observed with a short 
      equilibration time. Analysis of the binding data reveals a possible cooperative 
      binding process in solution. The nature of the fluorophore clearly influences the 
      sensitivity of the analysis more than the tether length used for the dye 
      conjugation. The local fluorophore motion is seen to influence the sensitivity of 
      the FP probe significantly. Texas Red is seen to be relatively more sensitive for 
      this approach and has apparently favorable dye-DNA interactions, and a limit of 
      detection of 350 pM was obtained. Significant temperature dependence of the FP 
      responses has been observed in this work. Ionic composition of the binding buffer 
      also influences the assay sensitivity. The results confirm the promise and 
      potential of similar homogeneous assays for aptamer-based bioanalysis.
FAU - Gokulrangan, Giridharan
AU  - Gokulrangan G
AD  - Department of Chemistry, University of Kansas, Lawrence, Kansas 66045, USA.
FAU - Unruh, Jay R
AU  - Unruh JR
FAU - Holub, Douglas F
AU  - Holub DF
FAU - Ingram, Brian
AU  - Ingram B
FAU - Johnson, Carey K
AU  - Johnson CK
FAU - Wilson, George S
AU  - Wilson GS
LA  - eng
GR  - 5 T32 GM08545-09/GM/NIGMS NIH HHS/United States
PT  - Comparative Study
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Anal Chem
JT  - Analytical chemistry
JID - 0370536
RN  - 0 (Aptamers, Nucleotide)
RN  - 0 (Fluorescent Dyes)
RN  - 0 (Xanthenes)
RN  - 37341-29-0 (Immunoglobulin E)
RN  - 82354-19-6 (Texas red)
RN  - TPY09G7XIR (Fluorescein)
SB  - IM
MH  - Aptamers, Nucleotide/*chemistry/*metabolism
MH  - Fluorescein/chemistry
MH  - Fluorescence Polarization/*methods
MH  - Fluorescent Dyes/chemistry
MH  - Immunoglobulin E/*analysis/metabolism
MH  - Sensitivity and Specificity
MH  - Temperature
MH  - Xanthenes/chemistry
EDAT- 2005/04/02 09:00
MHDA- 2007/02/14 09:00
CRDT- 2005/04/02 09:00
PHST- 2005/04/02 09:00 [pubmed]
PHST- 2007/02/14 09:00 [medline]
PHST- 2005/04/02 09:00 [entrez]
AID - 10.1021/ac0483926 [doi]
PST - ppublish
SO  - Anal Chem. 2005 Apr 1;77(7):1963-70. doi: 10.1021/ac0483926.