HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

J. Biol. Chem., Vol. 280, Issue 40, 34193-34201, October 7, 2005

This Article Full Text Full Text (PDF) All Versions of this Article: 280/40/34193    most recent M504772200v1 Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Fitter, S. Articles by James, R. Search for Related Content PubMed PubMed Citation Articles by Fitter, S. Articles by James, R. Social Bookmarking                      What's this?

Deconvolution of a Complex Target Using DNA Aptamers^*

From the Department of Gastroenterology and Hepatology, Flinders Medical Centre, Adelaide, South Australia 5042

In vitro selection of single-stranded nucleic acid aptamers from large random sequence libraries is now a straightforward process particularly when screening with a single target molecule. These libraries contain considerable shape diversity as evident by the successful isolation of aptamers that bind with high affinity and specificity to chemically diverse targets. We propose that aptamer libraries contain sufficient shape diversity to allow deconvolution of a complex mixture of targets. Using unfractionated human plasma as our experimental model, we aim to develop methods to obtain aptamers against as many proteins as possible. To begin, it is critical that we understand how aptamer populations change with increasing rounds of in vitro selection when using complex mixtures. Our results show that sequence representation in the selected population changes dramatically with increasing rounds of selection. Certain aptamer families were apparent after only three selection rounds. Two additional cycles saw a decline in the relative abundance of these families and the emergence of yet another family that accounted for more than 60% of sequences in the pool. To overcome this population convergence, an aptamer-based target depletion method was developed, and the library screen was repeated. The previous dominant family effectively disappeared from the selected populations but was replaced by other aptamer families. Insights gained from these initial experiments are now being applied in the creation of second generation plasma protein screens and also to the analysis of other complex biological targets.

Received for publication, May 2, 2005 , and in revised form, July 7, 2005.

^* This work was supported in part by a grant from Flinders Technologies Pty. Ltd. with additional support provided from the South Australia Government and an Australian Industry Biotechnology Innovation Fund grant. Continuing support is provided by Aptamer Technologies PTY LTD. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

¹ Recipient of an Australian Postgraduate Award scholarship.

² To whom correspondence should be addressed: Dept. of Gastroenterology and Hepatology, Flinders Medical Centre, Bedford Park, South Australia, Australia, 5042. Tel.: 61-8-8204-5011; Fax: 61-8-8204-3943; E-mail: rob.james{at}flinders.edu.au.

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				K.-T. Guo, R. SchAfer, A. Paul, A. Gerber, G. Ziemer, and H. P. Wendel A New Technique for the Isolation and Surface Immobilization of Mesenchymal Stem Cells from Whole Bone Marrow Using High-Specific DNA Aptamers Stem Cells, October 1, 2006; 24(10): 2220 - 2231. [Abstract] [Full Text] [PDF]