First report of the use of a saxitoxin-protein conjugate to develop a DNA aptamer to a small molecule toxin

被引:79
作者
Handy, Sara M. [1 ]
Yakes, Betsy Jean [1 ]
DeGrasse, Jeffrey A. [1 ]
Campbell, Katrina [2 ]
Elliott, Christopher T. [2 ]
Kanyuck, Kelsey M. [3 ]
DeGrasse, Stacey L. [1 ]
机构
[1] US FDA, Ctr Food Safety & Appl Nutr, College Pk, MD 20740 USA
[2] Queens Univ Belfast, IAFLU, Sch Biol Sci, Belfast BT9 5AG, Antrim, North Ireland
[3] Univ Maryland, JIFSAN, College Pk, MD 20742 USA
关键词
Aptamer; Saxitoxin; Marine toxin; Paralytic shellfish toxins; Surface plasmon resonance; SELECTION; LIGANDS; SELEX; ACID; ANTIBODIES; ENRICHMENT; EVOLUTION; BINDING;
D O I
10.1016/j.toxicon.2012.10.015
中图分类号
R9 [药学];
学科分类号
1007 ;
摘要
Saxitoxin (STX) is a low molecular weight neurotoxin mainly produced by certain marine dinoflagellates that, along with its family of similarly related paralytic shellfish toxins, may cause the potentially fatal intoxication known as paralytic shellfish poisoning. Illness and fatality rates are low due to the effective monitoring programs that determine when toxins exceed the established regulatory action level and effectuate shellfish harvesting closures accordingly. Such monitoring programs rely on the ability to rapidly screen large volumes of samples. Many of the screening assays currently available employ antibodies or live animals. This research focused on developing an analytical recognition element that would eliminate the challenges associated with the limited availability of antibodies and the use of animals. Here we report the discovery of a DNA aptamer that targets STX. Concentration-dependent and selective binding of the aptamer to STX was determined using a surface plasmon resonance sensor. Not only does this work represent the first reported aptamer to SIX, but also the first aptamer to any marine biotoxin. A novel strategy of using a toxin protein conjugate for DNA aptamer selection was successfully implemented to overcome the challenges associated with aptamer selection to small molecules. Taking advantage of such an approach could lead to increased diversity and accessibility of aptamers to low molecular weight toxins, which could then be incorporated as analytical recognition elements in diagnostic assays for foodborne toxin detection. The selected STX aptamer sequence is provided here, making it available to any investigator for use in assay development for the detection of SIX. Published by Elsevier Ltd.
引用
收藏
页码:30 / 37
页数:8
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