Skip to main content
U.S. flag

An official website of the United States government

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you’ve safely connected to the .gov website. Share sensitive information only on official, secure websites.

Development of Aptamer-Based Affinity Assays Using Temperature Gradient Focusing: Minimization of the Limit of Detection

Published

Author(s)

Matt S. Munson, John Meacham, David J. Ross, Laurie E. Locascio

Abstract

A method is described for an aptamer-based affinity assay using a combination of two non-conventional techniques, temperature gradient focusing (TGF) and field amplified continuous sample injection TGF (FACSI-TGF), with fluorescence detection. Human immunodeficiency virus reverse transcriptase (HIVRT) is used as the protein target for the assay. The TGF and FACSI-TGF assays are compared to similar results obtained with conventional capillary electrophoresis (CE). A range of starting aptamer concentrations are used to determine the optimal limit of detection (LOD) for HIVRT using each approach. The results indicate that the detection limits for HIVRT obtained with TGF and FACSI-TGF are comparable to or even lower than the detection limits obtained with conventional CE in spite of the inferior detector used for the TGF and FACSI-TGF assays (arc lamp and low cost CCD for TGF vs. LIF with PMT for CE). It is hypothesized that this is due to the greater reproducibility of the TGF and FACSI-TGF techniques since they do not employ a defined sample injection. The lowest detection limit achieved with the new aptamer assay approach is more than two orders of magnitude lower than that reported for a similar CE-based aptamer assay for the same target.
Citation
Electrophoresis
Volume
29
Issue
16

Citation

Munson, M. , Meacham, J. , Ross, D. and Locascio, L. (2008), Development of Aptamer-Based Affinity Assays Using Temperature Gradient Focusing: Minimization of the Limit of Detection, Electrophoresis, [online], https://tsapps.nist.gov/publication/get_pdf.cfm?pub_id=832288 (Accessed April 22, 2024)
Created August 1, 2008, Updated February 19, 2017