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Screening and initial binding assessment of fumonisin b(1) aptamers.

McKeague M, Bradley CR, De Girolamo A, Visconti A, Miller JD, Derosa MC - Int J Mol Sci (2010)

Bottom Line: The use of aptamers offers a good alternative to the use of antibodies in fumonisin cleanup and detection due to lower costs and improved stability.Six unique sequences were obtained, each showing improved binding to fumonisin B(1) compared to controls.Sequence FB(1) 39 binds to fumonisin with a dissociation constant of 100 ± 30 nM and shows potential for use in fumonisin biosensors and solid phase extraction columns.

View Article: PubMed Central - PubMed

Affiliation: Chemistry Department, Carleton University, 1125 Colonel By Drive, Ottawa, ON, Canada K1S 5B6; E-Mails: mmckeagu@connect.carleton.ca (M.M.); cbradle4@connect.carleton.ca (C.R.B.); david_miller@carleton.ca (J.D.M.).

ABSTRACT
Fumonisins are mycotoxins produced by Fusarium verticillioides and F. proliferatum, fungi that are ubiquitous in corn (maize). Insect damage and some other environmental conditions result in the accumulation of fumonisins in corn-based products worldwide. Current methods of fumonisin detection rely on the use of immunoaffinity columns and high-performance liquid chromatography (HPLC). The use of aptamers offers a good alternative to the use of antibodies in fumonisin cleanup and detection due to lower costs and improved stability. Aptamers are single-stranded oligonucleotides that are selected using Systematic Evolution of Ligands by EXponential enrichment (SELEX) for their ability to bind to targets with high affinity and specificity. Sequences obtained after 18 rounds of SELEX were screened for their ability to bind to fumonisin B(1). Six unique sequences were obtained, each showing improved binding to fumonisin B(1) compared to controls. Sequence FB(1) 39 binds to fumonisin with a dissociation constant of 100 ± 30 nM and shows potential for use in fumonisin biosensors and solid phase extraction columns.

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Related in: MedlinePlus

Binding curves for the aptamer sequence FB1 39. Three trials were performed to obtain saturation curves used to determine the dissociation constant (KD). Bound fluorescein-labeled aptamer was measured by fluorescence (excitation at 490 nm, emission at 518 nm).
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f5-ijms-11-04864: Binding curves for the aptamer sequence FB1 39. Three trials were performed to obtain saturation curves used to determine the dissociation constant (KD). Bound fluorescein-labeled aptamer was measured by fluorescence (excitation at 490 nm, emission at 518 nm).

Mentions: The binding affinity of sequence FB1 39 was investigated in more detail and was found to interact with immobilized FB1 with a dissociation constant, KD, of 100 ± 30 nM (see Figure 5). Therefore, this full length aptamer clone displays a high affinity for this relatively small molecular target. Comparable binding of the aptamer to the target when in solution is expected and will be determined in our future work. Further analysis will also attempt to identify the minimal target-binding sequence to shorten the aptamer and improve binding while lowering the aptamer’s production cost.


Screening and initial binding assessment of fumonisin b(1) aptamers.

McKeague M, Bradley CR, De Girolamo A, Visconti A, Miller JD, Derosa MC - Int J Mol Sci (2010)

Binding curves for the aptamer sequence FB1 39. Three trials were performed to obtain saturation curves used to determine the dissociation constant (KD). Bound fluorescein-labeled aptamer was measured by fluorescence (excitation at 490 nm, emission at 518 nm).
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC3100853&req=5

f5-ijms-11-04864: Binding curves for the aptamer sequence FB1 39. Three trials were performed to obtain saturation curves used to determine the dissociation constant (KD). Bound fluorescein-labeled aptamer was measured by fluorescence (excitation at 490 nm, emission at 518 nm).
Mentions: The binding affinity of sequence FB1 39 was investigated in more detail and was found to interact with immobilized FB1 with a dissociation constant, KD, of 100 ± 30 nM (see Figure 5). Therefore, this full length aptamer clone displays a high affinity for this relatively small molecular target. Comparable binding of the aptamer to the target when in solution is expected and will be determined in our future work. Further analysis will also attempt to identify the minimal target-binding sequence to shorten the aptamer and improve binding while lowering the aptamer’s production cost.

Bottom Line: The use of aptamers offers a good alternative to the use of antibodies in fumonisin cleanup and detection due to lower costs and improved stability.Six unique sequences were obtained, each showing improved binding to fumonisin B(1) compared to controls.Sequence FB(1) 39 binds to fumonisin with a dissociation constant of 100 ± 30 nM and shows potential for use in fumonisin biosensors and solid phase extraction columns.

View Article: PubMed Central - PubMed

Affiliation: Chemistry Department, Carleton University, 1125 Colonel By Drive, Ottawa, ON, Canada K1S 5B6; E-Mails: mmckeagu@connect.carleton.ca (M.M.); cbradle4@connect.carleton.ca (C.R.B.); david_miller@carleton.ca (J.D.M.).

ABSTRACT
Fumonisins are mycotoxins produced by Fusarium verticillioides and F. proliferatum, fungi that are ubiquitous in corn (maize). Insect damage and some other environmental conditions result in the accumulation of fumonisins in corn-based products worldwide. Current methods of fumonisin detection rely on the use of immunoaffinity columns and high-performance liquid chromatography (HPLC). The use of aptamers offers a good alternative to the use of antibodies in fumonisin cleanup and detection due to lower costs and improved stability. Aptamers are single-stranded oligonucleotides that are selected using Systematic Evolution of Ligands by EXponential enrichment (SELEX) for their ability to bind to targets with high affinity and specificity. Sequences obtained after 18 rounds of SELEX were screened for their ability to bind to fumonisin B(1). Six unique sequences were obtained, each showing improved binding to fumonisin B(1) compared to controls. Sequence FB(1) 39 binds to fumonisin with a dissociation constant of 100 ± 30 nM and shows potential for use in fumonisin biosensors and solid phase extraction columns.

Show MeSH
Related in: MedlinePlus