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Study of Immobilization Procedure on Silver Nanolayers and Detection of Estrone with Diverged Beam Surface Plasmon Resonance (SPR) Imaging.

Karabchevsky A, Tsapovsky L, Marks RS, Abdulhalim I - Biosensors (Basel) (2013)

Bottom Line: The signals, during each step of the protocol, were analyzed with a refractometer based on the surface plasmon resonance (SPR) effect and luminescence techniques.To demonstrate specific biosensing, rabbit anti-estrone polyclonal immunoglobulin G (IgG) antibody was immobilized through a linker on 47 nm silver layer deposited on SF11 glass.At the final stage, the representative endocrine disruptor-estrone-was attached and detected in deionized water with a diverging beam SPR imaging sensor.

View Article: PubMed Central - PubMed

Affiliation: Electro-Optic Engineering Unit and Ilse Katz Institute for Nanoscale Science and Technology, Ben-Gurion University of the Negev, Beer-Sheva 84105, Israel; E-Mail: abdulhlm@bgu.ac.il.

ABSTRACT
An immobilization protocol was developed to attach receptors on smooth silver thin films. Dense and packed 11-mercaptoundecanoic acid (11-MUA) was used to avoid uncontrolled sulfidization and harmful oxidation of silver nanolayers. N,N'-dicyclohexylcarbodiimide (DCC) and N-hydroxysuccinimide (NHS) were added to make the silver surfaces reactive. A comparative study was carried out with different immersion times of silver samples in 11-MUA solutions with different concentrations to find the optimum conditions for immobilization. The signals, during each step of the protocol, were analyzed with a refractometer based on the surface plasmon resonance (SPR) effect and luminescence techniques. Molecular interactions at the surfaces between the probe and target at the surface nanolayer shift the SPR signal, thus indicating the presence of the substance. To demonstrate specific biosensing, rabbit anti-estrone polyclonal immunoglobulin G (IgG) antibody was immobilized through a linker on 47 nm silver layer deposited on SF11 glass. At the final stage, the representative endocrine disruptor-estrone-was attached and detected in deionized water with a diverging beam SPR imaging sensor.

No MeSH data available.


Related in: MedlinePlus

Reflected images from DBSPRI sensor, top: 68 nm Ag on BK7 glass after incubation in (a) 10 mM 11-MUA for 24 h and (b) after incubation in 10 mM 11-MUA for 24 h immobilized rabbit anti-estrone polyclonal IgG, while adding NHS + DCC prior to the immobilization; bottom: analyzed images in Radon space.
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biosensors-03-00157-f006: Reflected images from DBSPRI sensor, top: 68 nm Ag on BK7 glass after incubation in (a) 10 mM 11-MUA for 24 h and (b) after incubation in 10 mM 11-MUA for 24 h immobilized rabbit anti-estrone polyclonal IgG, while adding NHS + DCC prior to the immobilization; bottom: analyzed images in Radon space.

Mentions: First, we validated the rabbit anti-estrone polyclonal IgG immobilization and attachment of the antigen—estrone—to it (Figure 5) with standard KR configuration, which is schematically shown in Figure 2. As is expected, the SPR angle increases as more molecules attached to the silver surface. After that, we demonstrated specific sensing of estrone with a DBSPRI sensor and analyzed the signals with a Radon transform-based algorithm, which was detailed in our previous work [24]. Figure 6 shows reflected signals from the DBSPRI sensor (depicted in Figure 3), while rabbit anti-estrone polyclonal IgG was immobilized on the silver surface. On top of Figure 6 is shown captured image, while 68 nm Ag on BK7 glass was incubated in (a) 10 mM 11-MUA for 24 h and (b) rabbit anti-estrone polyclonal IgG was immobilized, while adding NHS+DCC prior to the immobilization; down: analyzed images in Radon space.


Study of Immobilization Procedure on Silver Nanolayers and Detection of Estrone with Diverged Beam Surface Plasmon Resonance (SPR) Imaging.

Karabchevsky A, Tsapovsky L, Marks RS, Abdulhalim I - Biosensors (Basel) (2013)

Reflected images from DBSPRI sensor, top: 68 nm Ag on BK7 glass after incubation in (a) 10 mM 11-MUA for 24 h and (b) after incubation in 10 mM 11-MUA for 24 h immobilized rabbit anti-estrone polyclonal IgG, while adding NHS + DCC prior to the immobilization; bottom: analyzed images in Radon space.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4263594&req=5

biosensors-03-00157-f006: Reflected images from DBSPRI sensor, top: 68 nm Ag on BK7 glass after incubation in (a) 10 mM 11-MUA for 24 h and (b) after incubation in 10 mM 11-MUA for 24 h immobilized rabbit anti-estrone polyclonal IgG, while adding NHS + DCC prior to the immobilization; bottom: analyzed images in Radon space.
Mentions: First, we validated the rabbit anti-estrone polyclonal IgG immobilization and attachment of the antigen—estrone—to it (Figure 5) with standard KR configuration, which is schematically shown in Figure 2. As is expected, the SPR angle increases as more molecules attached to the silver surface. After that, we demonstrated specific sensing of estrone with a DBSPRI sensor and analyzed the signals with a Radon transform-based algorithm, which was detailed in our previous work [24]. Figure 6 shows reflected signals from the DBSPRI sensor (depicted in Figure 3), while rabbit anti-estrone polyclonal IgG was immobilized on the silver surface. On top of Figure 6 is shown captured image, while 68 nm Ag on BK7 glass was incubated in (a) 10 mM 11-MUA for 24 h and (b) rabbit anti-estrone polyclonal IgG was immobilized, while adding NHS+DCC prior to the immobilization; down: analyzed images in Radon space.

Bottom Line: The signals, during each step of the protocol, were analyzed with a refractometer based on the surface plasmon resonance (SPR) effect and luminescence techniques.To demonstrate specific biosensing, rabbit anti-estrone polyclonal immunoglobulin G (IgG) antibody was immobilized through a linker on 47 nm silver layer deposited on SF11 glass.At the final stage, the representative endocrine disruptor-estrone-was attached and detected in deionized water with a diverging beam SPR imaging sensor.

View Article: PubMed Central - PubMed

Affiliation: Electro-Optic Engineering Unit and Ilse Katz Institute for Nanoscale Science and Technology, Ben-Gurion University of the Negev, Beer-Sheva 84105, Israel; E-Mail: abdulhlm@bgu.ac.il.

ABSTRACT
An immobilization protocol was developed to attach receptors on smooth silver thin films. Dense and packed 11-mercaptoundecanoic acid (11-MUA) was used to avoid uncontrolled sulfidization and harmful oxidation of silver nanolayers. N,N'-dicyclohexylcarbodiimide (DCC) and N-hydroxysuccinimide (NHS) were added to make the silver surfaces reactive. A comparative study was carried out with different immersion times of silver samples in 11-MUA solutions with different concentrations to find the optimum conditions for immobilization. The signals, during each step of the protocol, were analyzed with a refractometer based on the surface plasmon resonance (SPR) effect and luminescence techniques. Molecular interactions at the surfaces between the probe and target at the surface nanolayer shift the SPR signal, thus indicating the presence of the substance. To demonstrate specific biosensing, rabbit anti-estrone polyclonal immunoglobulin G (IgG) antibody was immobilized through a linker on 47 nm silver layer deposited on SF11 glass. At the final stage, the representative endocrine disruptor-estrone-was attached and detected in deionized water with a diverging beam SPR imaging sensor.

No MeSH data available.


Related in: MedlinePlus