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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

Schematic of the immobilization steps: (a) incubation in 11-MUA, (b) adding N-hydroxysuccinimide (NHS) + 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide (EDC) (or N,N'-dicyclohexylcarbodiimide (DCC)), (c) immobilization of antibody and (d) attachment of antigen.
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biosensors-03-00157-f001: Schematic of the immobilization steps: (a) incubation in 11-MUA, (b) adding N-hydroxysuccinimide (NHS) + 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide (EDC) (or N,N'-dicyclohexylcarbodiimide (DCC)), (c) immobilization of antibody and (d) attachment of antigen.

Mentions: We analyzed the refractive index (RI) changes at the end of each step of the immobilization protocol. The steps of the immobilization procedure are schematically shown in Figure 1. They included: (1) samples of smooth silver layers after deposition on glass, (2) incubation in 11-MUA of different concentrations and immersion times, (3) adding the NHS + EDC or NHS + DCC to make the silver surfaces reactive, (4) immobilization of the antibody and (5) attachment of the antigen.


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)

Schematic of the immobilization steps: (a) incubation in 11-MUA, (b) adding N-hydroxysuccinimide (NHS) + 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide (EDC) (or N,N'-dicyclohexylcarbodiimide (DCC)), (c) immobilization of antibody and (d) attachment of antigen.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

biosensors-03-00157-f001: Schematic of the immobilization steps: (a) incubation in 11-MUA, (b) adding N-hydroxysuccinimide (NHS) + 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide (EDC) (or N,N'-dicyclohexylcarbodiimide (DCC)), (c) immobilization of antibody and (d) attachment of antigen.
Mentions: We analyzed the refractive index (RI) changes at the end of each step of the immobilization protocol. The steps of the immobilization procedure are schematically shown in Figure 1. They included: (1) samples of smooth silver layers after deposition on glass, (2) incubation in 11-MUA of different concentrations and immersion times, (3) adding the NHS + EDC or NHS + DCC to make the silver surfaces reactive, (4) immobilization of the antibody and (5) attachment of the antigen.

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