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Fabrication of surface plasmon resonance sensor surface with control of the non-specific adsorption and affinity for the detection of 2,4,6-trinitrotoluene using an antifouling copolymer.

Yatabe R, Onodera T, Toko K - Front Bioeng Biotechnol (2014)

Bottom Line: We fabricated a surface plasmon resonance (SPR) sensor using a hydrophilic polymer for the highly sensitive detection of 2,4,6-trinitrotoluene (TNT).The detection of TNT was carried out by displacement assay with the SPR measurement.In displacement assay, the affinity between anti-TNT antibody and the sensor surface, affects to the sensitivity.

View Article: PubMed Central - PubMed

Affiliation: Research and Development Center for Taste and Odor Sensing, Kyushu University , Fukuoka , Japan.

ABSTRACT
We fabricated a surface plasmon resonance (SPR) sensor using a hydrophilic polymer for the highly sensitive detection of 2,4,6-trinitrotoluene (TNT). The hydrophilic polymer was made from mono-2-(methacryloyloxy)ethylsuccinate (MES) and 2-hydroxyethylmethacrylate (HEMA) by surface-initiated atom transfer radical polymerization. The detection of TNT was carried out by displacement assay with the SPR measurement. In displacement assay, the affinity between anti-TNT antibody and the sensor surface, affects to the sensitivity. In the SPR measurement, non-specific adsorption should be controlled because SPR sensor cannot discriminate between specific and non-specific adsorption. Therefore, the affinity and non-specific adsorption were controlled by changing the ratio of HEMA to MES. A detection limit of 0.4 ng/ml (ppb) for TNT was achieved using a sensor surface with the lowest affinity without non-specific adsorption.

No MeSH data available.


Related in: MedlinePlus

Adsorption of lysozyme, BSA, and anti-TNT antibody.
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Figure 2: Adsorption of lysozyme, BSA, and anti-TNT antibody.

Mentions: The amount of adsorption on the surface was measured by SPR when 25 ppm anti-TNT antibody, 1000 ppm lysozyme, or 1000 ppm BSA was allowed to flow on the surface for 2 min. The antibody can adsorb on the surface by specific/non-specific adsorption. Lysozyme, which is positively charged, can adsorb on the surface by electrostatic interaction if the surface is negatively charged. BSA, which is negatively charged, can adsorb on the surface if the surface is positively charged. In addition, lysozyme and BSA can adsorb on the surface by a hydrophobic interaction if the surface has hydrophobicity. Figure 2 shows the results of SPR measurement of the surface with various ratios of MES to HEMA. First, the surface was not positively charged, and did not have hydrophobicity because BSA was not adsorbed on the surface under any of the conditions. Second, the amount of lysozyme adsorbed decreased with increasing amount of HEMA. When the amount of HEMA was high, negligible amount of lysozyme were able to adsorb on the surface by electrostatic interaction because the negative charge on the surface became weaker. A surface with low non-specific adsorption was obtained at MES:HEMA ratios of 1:10, 1:100, and 1:1000. Third, at MES:HEMA ratios of 1:0, 1:1, 1:2, 1:5, and 1:10, the amount of antibody adsorbed increased with the amount of HEMA. It is suggested that the antibody we used was negatively charged because of this result and that in our previous study: the amount of antibody adsorbed was large when the polymer surface was positively charged (Yatabe et al., 2013b). Adsorption of the antibody, which can be negatively charged, on the surface was negligible because the surface was negatively charged when the amount of HEMA was small. Next, the amount of antibody absorbed became maximum when the MES:HEMA ratio was 1:10 or 1:100. The amount of antibody absorbed was smaller for the MES:HEMA ratio of 1:1000 than for the ratio of 1:100. For the MES:HEMA ratio of 1:1000, the amount of MES was the smallest among all of the conditions. The binding site of the antibody was DNP-Hdrz, which was immobilized on the carboxyl group of MES. The number of binding sites decreased when the amount of MES was small. It was difficult for the antibody to adsorb on the surface because the amount of immobilized DNP-Hdrz was small. In conclusion, a sensor surface with low non-specific adsorption and capable of immunoreaction was fabricated by mixing MES and HEMA.


Fabrication of surface plasmon resonance sensor surface with control of the non-specific adsorption and affinity for the detection of 2,4,6-trinitrotoluene using an antifouling copolymer.

Yatabe R, Onodera T, Toko K - Front Bioeng Biotechnol (2014)

Adsorption of lysozyme, BSA, and anti-TNT antibody.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4126440&req=5

Figure 2: Adsorption of lysozyme, BSA, and anti-TNT antibody.
Mentions: The amount of adsorption on the surface was measured by SPR when 25 ppm anti-TNT antibody, 1000 ppm lysozyme, or 1000 ppm BSA was allowed to flow on the surface for 2 min. The antibody can adsorb on the surface by specific/non-specific adsorption. Lysozyme, which is positively charged, can adsorb on the surface by electrostatic interaction if the surface is negatively charged. BSA, which is negatively charged, can adsorb on the surface if the surface is positively charged. In addition, lysozyme and BSA can adsorb on the surface by a hydrophobic interaction if the surface has hydrophobicity. Figure 2 shows the results of SPR measurement of the surface with various ratios of MES to HEMA. First, the surface was not positively charged, and did not have hydrophobicity because BSA was not adsorbed on the surface under any of the conditions. Second, the amount of lysozyme adsorbed decreased with increasing amount of HEMA. When the amount of HEMA was high, negligible amount of lysozyme were able to adsorb on the surface by electrostatic interaction because the negative charge on the surface became weaker. A surface with low non-specific adsorption was obtained at MES:HEMA ratios of 1:10, 1:100, and 1:1000. Third, at MES:HEMA ratios of 1:0, 1:1, 1:2, 1:5, and 1:10, the amount of antibody adsorbed increased with the amount of HEMA. It is suggested that the antibody we used was negatively charged because of this result and that in our previous study: the amount of antibody adsorbed was large when the polymer surface was positively charged (Yatabe et al., 2013b). Adsorption of the antibody, which can be negatively charged, on the surface was negligible because the surface was negatively charged when the amount of HEMA was small. Next, the amount of antibody absorbed became maximum when the MES:HEMA ratio was 1:10 or 1:100. The amount of antibody absorbed was smaller for the MES:HEMA ratio of 1:1000 than for the ratio of 1:100. For the MES:HEMA ratio of 1:1000, the amount of MES was the smallest among all of the conditions. The binding site of the antibody was DNP-Hdrz, which was immobilized on the carboxyl group of MES. The number of binding sites decreased when the amount of MES was small. It was difficult for the antibody to adsorb on the surface because the amount of immobilized DNP-Hdrz was small. In conclusion, a sensor surface with low non-specific adsorption and capable of immunoreaction was fabricated by mixing MES and HEMA.

Bottom Line: We fabricated a surface plasmon resonance (SPR) sensor using a hydrophilic polymer for the highly sensitive detection of 2,4,6-trinitrotoluene (TNT).The detection of TNT was carried out by displacement assay with the SPR measurement.In displacement assay, the affinity between anti-TNT antibody and the sensor surface, affects to the sensitivity.

View Article: PubMed Central - PubMed

Affiliation: Research and Development Center for Taste and Odor Sensing, Kyushu University , Fukuoka , Japan.

ABSTRACT
We fabricated a surface plasmon resonance (SPR) sensor using a hydrophilic polymer for the highly sensitive detection of 2,4,6-trinitrotoluene (TNT). The hydrophilic polymer was made from mono-2-(methacryloyloxy)ethylsuccinate (MES) and 2-hydroxyethylmethacrylate (HEMA) by surface-initiated atom transfer radical polymerization. The detection of TNT was carried out by displacement assay with the SPR measurement. In displacement assay, the affinity between anti-TNT antibody and the sensor surface, affects to the sensitivity. In the SPR measurement, non-specific adsorption should be controlled because SPR sensor cannot discriminate between specific and non-specific adsorption. Therefore, the affinity and non-specific adsorption were controlled by changing the ratio of HEMA to MES. A detection limit of 0.4 ng/ml (ppb) for TNT was achieved using a sensor surface with the lowest affinity without non-specific adsorption.

No MeSH data available.


Related in: MedlinePlus