PMID- 23688685
OWN - NLM
STAT- MEDLINE
DCOM- 20131104
LR  - 20191210
IS  - 1873-3778 (Electronic)
IS  - 0021-9673 (Linking)
VI  - 1295
DP  - 2013 Jun 21
TI  - Aptamer-protein binding detected by asymmetric flow field flow fractionation.
PG  - 107-13
LID - S0021-9673(13)00689-4 [pii]
LID - 10.1016/j.chroma.2013.04.063 [doi]
AB  - Asymmetric flow field flow fractionation (AF4) should be suitable for the study 
      of aptamer-target binding, because its gentle separation would impose little 
      disturbance to the complex structure, and it can use carrier solutions with high 
      salt concentrations to provide the most optimal interaction environment to the 
      complex. However, no report has been found for such applications. Herein, we 
      investigated the utility of AF4 as an effective tool for detection of the 
      aptamer-protein complex. With the model system of human immunoglobulin E (IgE) 
      and its aptamer, impacts on aptamer binding from the incubation and AF4 carrier 
      solutions, as well as the flow conditions used during the sample focusing step, 
      were studied. We found that the composition of the carrier solution, in 
      particular, the presence of Mg(2+), strongly influenced the complex's integrity 
      in AF4. Also, the focusing conditions during sample injection in AF4 affected the 
      binding equilibrium. Our findings highlight the necessity of maintaining the 
      optimal binding environment during the time course of complex measurement; and 
      demonstrated the good compatibility of AF4 with salty buffers and its high 
      simplicity in conducting on-channel incubation. With its capability to carry out 
      size-based separation of analytes with a wide range of dimensions, AF4 can be 
      employed for detection of large proteins and even biological particles using 
      aptamers. AF4 is also valuable for study of aptamer-target binding under 
      different buffer environments for better understanding of the structure-function 
      relationship of aptamers.
CI  - Copyright (c) 2013 Elsevier B.V. All rights reserved.
FAU - Schachermeyer, Samantha
AU  - Schachermeyer S
AD  - Department of Chemistry, University of California, Riverside, CA 92521, USA.
FAU - Ashby, Jonathan
AU  - Ashby J
FAU - Zhong, Wenwan
AU  - Zhong W
LA  - eng
PT  - Evaluation Study
PT  - Journal Article
PT  - Research Support, U.S. Gov't, Non-P.H.S.
DEP - 20130428
PL  - Netherlands
TA  - J Chromatogr A
JT  - Journal of chromatography. A
JID - 9318488
RN  - 0 (Aptamers, Peptide)
RN  - 37341-29-0 (Immunoglobulin E)
SB  - IM
MH  - Aptamers, Peptide/*chemistry
MH  - Fractionation, Field Flow/instrumentation/*methods
MH  - Immunoglobulin E/*chemistry
MH  - Protein Binding
EDAT- 2013/05/22 06:00
MHDA- 2013/11/05 06:00
CRDT- 2013/05/22 06:00
PHST- 2013/02/23 00:00 [received]
PHST- 2013/04/18 00:00 [revised]
PHST- 2013/04/19 00:00 [accepted]
PHST- 2013/05/22 06:00 [entrez]
PHST- 2013/05/22 06:00 [pubmed]
PHST- 2013/11/05 06:00 [medline]
AID - S0021-9673(13)00689-4 [pii]
AID - 10.1016/j.chroma.2013.04.063 [doi]
PST - ppublish
SO  - J Chromatogr A. 2013 Jun 21;1295:107-13. doi: 10.1016/j.chroma.2013.04.063. Epub 
      2013 Apr 28.