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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

(a) Reflected images from DBSPRI sensor of 11-MUA + (NHS and DCC) after immobilization of estrone antibody and attached to it the estrone and after washing with PBS; (b) detection steps using the DBSPRI extraction algorithm [32]: blue—g(x), which is integrated original image; green—median filtered integrated image’ and red—median filtered image after background suppression. SPR line location is pointed by vertical lines; (c) reflected light, which was captured by the camera using the optical arrangement described in Figure 3, with extracted locations of the SPR signal, indicated by a solid line along the image.
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biosensors-03-00157-f007: (a) Reflected images from DBSPRI sensor of 11-MUA + (NHS and DCC) after immobilization of estrone antibody and attached to it the estrone and after washing with PBS; (b) detection steps using the DBSPRI extraction algorithm [32]: blue—g(x), which is integrated original image; green—median filtered integrated image’ and red—median filtered image after background suppression. SPR line location is pointed by vertical lines; (c) reflected light, which was captured by the camera using the optical arrangement described in Figure 3, with extracted locations of the SPR signal, indicated by a solid line along the image.

Mentions: Figure 7(a) shows the reflected image from the DBSPRI sensor described in Figure 3. Estrone antibody was immobilized and the antigen—estrone—was attached to it. Prior to the immobilization, the sample was incubated in 11-MUA for 24 h and NHS with DCC, which were added to make the surface reactive. Figure 7(b) shows detection steps using a multichannel extraction algorithm [32]: blue—g(x), which is integrated original image; green—median filtered integrated image; and red—median filtered image after background suppression. SPR line location is pointed by vertical lines. Figure 7(c) shows the extracted SPR line along the image.


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)

(a) Reflected images from DBSPRI sensor of 11-MUA + (NHS and DCC) after immobilization of estrone antibody and attached to it the estrone and after washing with PBS; (b) detection steps using the DBSPRI extraction algorithm [32]: blue—g(x), which is integrated original image; green—median filtered integrated image’ and red—median filtered image after background suppression. SPR line location is pointed by vertical lines; (c) reflected light, which was captured by the camera using the optical arrangement described in Figure 3, with extracted locations of the SPR signal, indicated by a solid line along the image.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

biosensors-03-00157-f007: (a) Reflected images from DBSPRI sensor of 11-MUA + (NHS and DCC) after immobilization of estrone antibody and attached to it the estrone and after washing with PBS; (b) detection steps using the DBSPRI extraction algorithm [32]: blue—g(x), which is integrated original image; green—median filtered integrated image’ and red—median filtered image after background suppression. SPR line location is pointed by vertical lines; (c) reflected light, which was captured by the camera using the optical arrangement described in Figure 3, with extracted locations of the SPR signal, indicated by a solid line along the image.
Mentions: Figure 7(a) shows the reflected image from the DBSPRI sensor described in Figure 3. Estrone antibody was immobilized and the antigen—estrone—was attached to it. Prior to the immobilization, the sample was incubated in 11-MUA for 24 h and NHS with DCC, which were added to make the surface reactive. Figure 7(b) shows detection steps using a multichannel extraction algorithm [32]: blue—g(x), which is integrated original image; green—median filtered integrated image; and red—median filtered image after background suppression. SPR line location is pointed by vertical lines. Figure 7(c) shows the extracted SPR line along the image.

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