Aptamer-Based Sensor Arrays for the Detection and Quantitation of Proteins

Department of Electrical & Computer Engineering, University of Texas at Austin, Austin, Texas, United States
Analytical Chemistry (Impact Factor: 5.64). 08/2004; 76(14):4066-75. DOI: 10.1021/ac049858n
Source: PubMed


Aptamer biosensors have been immobilized on beads, introduced into micromachined chips on the electronic tongue sensor array, and used for the detection and quantitation of proteins. Aptamer chips could detect proteins in both capture and sandwich assay formats. Unlike most protein-based arrays, the aptamer chips could be stripped and reused multiple times. The aptamer chips proved to be useful for screening aptamers from in vitro selection experiments and for sensitively quantitating the biothreat agent ricin.

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    • "3.1. Experimental principle of the aptamer-based EWA biosensor Aptamers exhibit high target-binding affinity because their binding function is largely dependent upon stable secondary structure interactions (Kirby et al., 2004; Zhou et al., 2010); however, it is challengeable to keep the restoration to the original DNA probe form after repeated uses (Liu and Tan, 1999). Inspired by the immunosensors undergoing the antibody–antigen recognition which feature with highly reusability of up to 300 times of regenerated test cycles (Rodriguez-Mozaz et al., 2005; Zhou et al., 2014), we describe here a novel aptamer-based EWA biosensing strategy as shown in Scheme 1. "
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    ABSTRACT: Although aptamer-based biosensors have attracted ever-increasing attentions and found potential applications in a wide range of areas, they usually adopted the assay protocol of immobilizing DNA probe (e.g., aptamer, aptamer-complementary oligonucleotides) on a solid sensing surface, making it critical and challengeable to keep the integration of nucleic acid surface during the regeneration and the restoration to its original DNA probe form after repeated uses. In order to address the issue, we report a novel aptamer-based biosensing strategy based on an evanescent wave all-fiber (EWA) platform. In a simple target capturing step using aptamer-functionalized magnetic microbeads, signal probes conjugated with streptavidin are released and further detected by a EWA biosensor via a facial dethiobiotin-streptavidin recognition. Apart from the inherent advantages of aptamer-based evanescent wave biosensors (e.g. target versatility, sensitivity, selectivity and portability), the proposed strategy exhibits a high stability and remarkable reusability over other aptasensors. Under the optimized conditions, the typical calibration curve obtained for Ochratoxin A has a detection limit of 3 nM with a linear response ranging from 6 nM to 500 nM. The dethiobiotin-streptavidin sensing surface instead of the traditional nucleic acid one can be reused for over 300 times without losing sensitivity.
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    • "Be‐ cause of their high thermostability when compared with antibodies, aptamers have potential applications in analytical devices, including biosensors, and as therapeutic agents [38]. Assays for protein identification and quantitation were developed and ap‐ plied to ricin detection [39] [40]. A multiplex aptamer microarray was generated by print‐ ing an anti-ricin RNA aptamer onto either streptavidin (SA)-or neutravidin (NA)-coated glass slides. "

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    • "c o m / l o c a t e / t ox i c on ricin in food and water. Conventional analytical methods including radioimmunoassay (Godal et al., 1981), enzymelinked immunosorbent assay (ELISA) (Poli et al., 1994), fluorescence-based fiber optic immunoassay (Narang et al., 1997) aptamer microarrays (Kirby et al., 2004), masssensitive biosensor (Tran et al., 2008), microelectrochemical biosensors (Guglielmo-Viret and Thullier, 2007) and electrochemiluminescent (ECL) assay (Garber and O'Brien, 2008) are effective for ricin detection. However these analytical methods are time-consuming and therefore not suitable for biodefense. "
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