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      Protein Detection with Aptamer Biosensors

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          Abstract

          Aptamers have been developed for different applications. Their use as new biological recognition elements in biosensors promises progress for fast and easy detection of proteins. This new generation of biosensor (aptasensors) will be more stable and well adapted to the conditions of real samples because of the specific properties of aptamers.

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          Systematic evolution of ligands by exponential enrichment: RNA ligands to bacteriophage T4 DNA polymerase.

          L Gold, C Tuerk (1990)
          High-affinity nucleic acid ligands for a protein were isolated by a procedure that depends on alternate cycles of ligand selection from pools of variant sequences and amplification of the bound species. Multiple rounds exponentially enrich the population for the highest affinity species that can be clonally isolated and characterized. In particular one eight-base region of an RNA that interacts with the T4 DNA polymerase was chosen and randomized. Two different sequences were selected by this procedure from the calculated pool of 65,536 species. One is the wild-type sequence found in the bacteriophage mRNA; one is varied from wild type at four positions. The binding constants of these two RNA's to T4 DNA polymerase are equivalent. These protocols with minimal modification can yield high-affinity ligands for any protein that binds nucleic acids as part of its function; high-affinity ligands could conceivably be developed for any target molecule.
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            SELEX--a (r)evolutionary method to generate high-affinity nucleic acid ligands.

            Regina Stoltenburg, Christine Reinemann, Beate Strehlitz (2007)
            SELEX stands for systematic evolution of ligands by exponential enrichment. This method, described primarily in 1990 [Ellington, A.D., Szostak, J.W., 1990. In vitro selection of RNA molecules that bind specific ligands. Nature 346, 818-822; Tuerk, C., Gold, L., 1990. Systematic evolution of ligands by exponential enrichment: RNA ligands to bacteriophage T4 DNA polymerase. Science 249, 505-510] aims at the development of aptamers, which are oligonucleotides (RNA or ssDNA) binding to their target with high selectivity and sensitivity because of their three-dimensional shape. Aptamers are all new ligands with a high affinity for considerably differing molecules ranging from large targets as proteins over peptides, complex molecules to drugs and organic small molecules or even metal ions. Aptamers are widely used, including medical and pharmaceutical basic research, drug development, diagnosis, and therapy. Analytical and separation tools bearing aptamers as molecular recognition and binding elements are another big field of application. Moreover, aptamers are used for the investigation of binding phenomena in proteomics. The SELEX method was modified over the years in different ways to become more efficient and less time consuming, to reach higher affinities of the aptamers selected and for automation of the process. This review is focused on the development of aptamers by use of SELEX and gives an overview about technologies, advantages, limitations, and applications of aptamers.
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              Aptamers: an emerging class of molecules that rival antibodies in diagnostics.

              S D Jayasena (1999)
              Antibodies, the most popular class of molecules providing molecular recognition needs for a wide range of applications, have been around for more than three decades. As a result, antibodies have made substantial contributions toward the advancement of diagnostic assays and have become indispensable in most diagnostic tests that are used routinely in clinics today. The development of the systematic evolution of ligands by exponential enrichment (SELEX) process, however, made possible the isolation of oligonucleotide sequences with the capacity to recognize virtually any class of target molecules with high affinity and specificity. These oligonucleotide sequences, referred to as "aptamers", are beginning to emerge as a class of molecules that rival antibodies in both therapeutic and diagnostic applications. Aptamers are different from antibodies, yet they mimic properties of antibodies in a variety of diagnostic formats. The demand for diagnostic assays to assist in the management of existing and emerging diseases is increasing, and aptamers could potentially fulfill molecular recognition needs in those assays. Compared with the bellwether antibody technology, aptamer research is still in its infancy, but it is progressing at a fast pace. The potential of aptamers may be realized in the near future in the form of aptamer-based diagnostic products in the market. In such products, aptamers may play a key role either in conjunction with, or in place of, antibodies. It is also likely that existing diagnostic formats may change according to the need to better harness the unique properties of aptamers.
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                Author and article information

                Journal
                Journal ID (nlm-ta): Sensors (Basel)
                Journal ID (iso-abbrev): Sensors (Basel)
                Title: Sensors (Basel, Switzerland)
                Publisher: Molecular Diversity Preservation International (MDPI)
                ISSN (Electronic): 1424-8220
                Publication date Collection: July 2008
                Publication date (Electronic): 23 July 2008
                Volume: 8
                Issue: 7
                Pages: 4296-4307
                Affiliations
                UFZ - Helmholtz Centre for Environmental Research, UBZ, Permoserstr. 15, 04318 Leipzig, Germany; E-Mails: nadia.nikolaus@ 123456ufz.de ; regina.stoltenburg@ 123456ufz.de
                Author notes
                [* ] Author to whom correspondence should be addressed; E-mail: beate.strehlitz@ 123456ufz.de ; Tel.: +49-341-235-1764; Fax: +49-341-235-451764
                Article
                Publisher ID: sensors-08-04296
                DOI: 10.3390/s8074296
                PMC ID: 3697175
                SO-VID: 87712d1d-04b9-4648-94ca-cbc2f9e2ae78
                Copyright © © 2008 by the authors; licensee Molecular Diversity Preservation International, Basel, Switzerland.
                License:

                This article is an open-access article distributed under the terms and conditions of the Creative Commons Attribution license ( http://creativecommons.org/licenses/by/3.0/).

                History
                Date received : 15 June 2008
                Date revision received : 07 July 2008
                Date accepted : 21 July 2008
                Categories
                Subject: Review

                ScienceOpen disciplines: Biomedical engineering
                Keywords: biosensor,protein,aptamer,selex
                Data availability:
                ScienceOpen disciplines: Biomedical engineering
                Keywords: biosensor, protein, aptamer, selex

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