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. 2008 Mar 15;18(6):1835-9.
doi: 10.1016/j.bmcl.2008.02.023. Epub 2008 Feb 13.

Combinatorial selection of a single stranded DNA thioaptamer targeting TGF-beta1 protein

Jonghoon Kang  1 , Myung Soog Lee, John A Copland 3rd, Bruce A Luxon, David G Gorenstein
Affiliations

Combinatorial selection of a single stranded DNA thioaptamer targeting TGF-beta1 protein

Jonghoon Kang et al. Bioorg Med Chem Lett. .

Abstract

A phosphorothioate single-stranded DNA aptamer (thioaptamer) targeting transforming growth factor-beta1 (TGF-beta1) was isolated by in-vitro combinatorial selection. The aptamer selection procedure was designed to modify the backbone of single-stranded DNA aptamers, where 5' of both A and C are phosphorothioates, since this provides enhanced nuclease resistance as well as higher affinity than that of a phosphate counterpart. The thioaptamer selected from a combinatorial library (5x10(14) sequences) binds to TGF-beta1 protein with an affinity of 90 nM. In this report, sequence, predicted secondary structure, and binding affinity of the selected thioaptamer (T18_1_3) are presented.

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Figures

Figure 1
Figure 1
Predicted secondary structure of T18_1_3. Initial and last nucleotides of the variable region are marked by arrows.
Figure 2
Figure 2
(A) EMSA of the initial thiolibrary (4 nM) with TGF-β1 protein. T and B represent top and bottom bands of the library, respectively. Data fitting to a Hill equation were shown in (B) where the circle and solid line is for the top band and the triangle and dashed line is for the bottom band.
Figure 3
Figure 3
(A) EMSA of a thioaptamer T18_1_3 (4 nM) with TGF-β1 protein. T, M, and B represent top, middle, and bottom bands of the thioaptamer, respectively. Data fitting to a Hill equation were shown in (B) where the circle and solid line is for the top band, the square and long dashed line is for the middle band, and the triangle and short dashed line is for the bottom band.
Figure 4
Figure 4
Multimerization of thioaptamer T18_1_3. (A) EMSA of DNA-DNA titration. (B) The ratios of band intensity of the top (triangle) and middle (circle) bands to the bottom band are plotted against the concentration of unlabeled T18_1_3.
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