Characterization of streptavidin binding to biotinylated, binary self-assembled thiol monolayers--influence of component ratio and solvent

Many biosensor applications are based on streptavidin (SA) binding to partially biotinylated self-assembled thiol monolayers (SAMs). In our study, binary SAMs on gold were prepared from solutions containing 16-mercapto-1-hexadecanol (thiol I) and N-(8-biotinyl-3,6-dioxa-octanamidyl)-16-mercaptohexad...

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Veröffentlicht in:Langmuir : the ACS journal of surfaces and colloids. - 1992. - 26(2010), 9 vom: 04. Mai, Seite 6386-93
1. Verfasser: Seifert, Michael (VerfasserIn)
Weitere Verfasser: Rinke, Matthias T, Galla, Hans-Joachim
Format: Online-Aufsatz
Sprache:English
Veröffentlicht: 2010
Zugriff auf das übergeordnete Werk:Langmuir : the ACS journal of surfaces and colloids
Schlagworte:Journal Article Research Support, Non-U.S. Gov't 16-mercaptohexadecanoic acid Hydroxides Immobilized Proteins Palmitic Acids Solvents Sulfhydryl Compounds Quartz 14808-60-7 mehr... Ethanol 3K9958V90M Chloroform 7V31YC746X Streptavidin 9013-20-1 hydroxide ion 9159UV381P
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100 1 |a Seifert, Michael  |e verfasserin  |4 aut 
245 1 0 |a Characterization of streptavidin binding to biotinylated, binary self-assembled thiol monolayers--influence of component ratio and solvent 
264 1 |c 2010 
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520 |a Many biosensor applications are based on streptavidin (SA) binding to partially biotinylated self-assembled thiol monolayers (SAMs). In our study, binary SAMs on gold were prepared from solutions containing 16-mercapto-1-hexadecanol (thiol I) and N-(8-biotinyl-3,6-dioxa-octanamidyl)-16-mercaptohexadecanamide (thiol II) in varying component ratios. Either chloroform or ethanol was used as solvent. After 24 h thiol incubation, SA was immobilized on the resulting SAMs using the strong SA-biotin interaction. The SA binding process was monitored by QCM-D (quartz crystal microbalance monitoring dissipation factor). It is shown that the Sauerbrey equation is valid to calculate the mass quantities of the immobilized SA layers. Under the chosen incubation conditions, marginal fractions of the biotinylated component II in chloroform ((n(I)/n(II))(solution) approximately = 1000) lead to SAMs which ensure a maximal SA binding quantity of m(Sauerbrey SA) approximately = 400 ng x cm(-2), being equivalent to a SA single-layer arrangement on the SAM surface. In case of incubations from ethanolic solutions, a complete SA layer formation needs significantly higher amounts of the biotinylated component II during SAM preparation ((n(I)/n(II))(solution) approximately = 50). X-ray photoelectron spectroscopy data show that the fraction of biotinylated thiol II in the SAM determines the amount of surface-bound SA. The SAM thiol ratio ((n(I)/n(II))(SAM)) not only depends on the corresponding component ratio in the incubation solution, but is also strongly influenced by the solvent. Using chloroform as solvent during SAM preparation significantly increased the fraction of biotinylated thiol II in the SAMs compared to ethanol 
650 4 |a Journal Article 
650 4 |a Research Support, Non-U.S. Gov't 
650 7 |a 16-mercaptohexadecanoic acid  |2 NLM 
650 7 |a Hydroxides  |2 NLM 
650 7 |a Immobilized Proteins  |2 NLM 
650 7 |a Palmitic Acids  |2 NLM 
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650 7 |a Sulfhydryl Compounds  |2 NLM 
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650 7 |a 14808-60-7  |2 NLM 
650 7 |a Ethanol  |2 NLM 
650 7 |a 3K9958V90M  |2 NLM 
650 7 |a Chloroform  |2 NLM 
650 7 |a 7V31YC746X  |2 NLM 
650 7 |a Streptavidin  |2 NLM 
650 7 |a 9013-20-1  |2 NLM 
650 7 |a hydroxide ion  |2 NLM 
650 7 |a 9159UV381P  |2 NLM 
700 1 |a Rinke, Matthias T  |e verfasserin  |4 aut 
700 1 |a Galla, Hans-Joachim  |e verfasserin  |4 aut 
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