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Biomolecular Interaction Analysis of Gestrinone-anti-Gestrinone Using Arrays of High Aspect Ratio SU-8 Nanopillars.

Ortega FJ, Bañuls MJ, Sanza FJ, Casquel R, Laguna MF, Holgado M, López-Romero D, Barrios CA, Maquieira Á, Puchades R - Biosensors (Basel) (2012)

Bottom Line: After gestrinone antigen immobilization on the BICELLs, the immunorecognition was performed.The cells were interrogated vertically by using micron spot size Fourier transform visible and IR spectrometry (FT-VIS-IR), and the dip wavenumber shift was monitored.The biosensing assay exhibited good reproducibility and sensitivity (LOD = 0.75 ng/mL).

View Article: PubMed Central - PubMed

Affiliation: Centro de Reconocimiento Molecular y Desarrollo Tecnológico, Departamento de Química, Universitat Politècnica de València, Camino de Vera s/n, Valencia 46022, Spain. fraorhi1@upvnet.upv.es.

ABSTRACT
In this paper, label-free biosensing for antibody screening by periodic lattices of high-aspect ratio SU-8 nano-pillars (BICELLs) is presented. As a demonstration, the determination of anti-gestrinone antibodies from whole rabbit serum is carried out, and for the first time, the dissociation constant (KD = 6 nM) of antigen-antibody recognition process is calculated using this sensing system. After gestrinone antigen immobilization on the BICELLs, the immunorecognition was performed. The cells were interrogated vertically by using micron spot size Fourier transform visible and IR spectrometry (FT-VIS-IR), and the dip wavenumber shift was monitored. The biosensing assay exhibited good reproducibility and sensitivity (LOD = 0.75 ng/mL).

No MeSH data available.


Related in: MedlinePlus

Dip shift for HRP-h-G immobilization and anti-gestrinone antibody recognition for the dip centered at 15,300 cm−1. Each measurement is the average of five replicates.
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biosensors-02-00291-f005: Dip shift for HRP-h-G immobilization and anti-gestrinone antibody recognition for the dip centered at 15,300 cm−1. Each measurement is the average of five replicates.

Mentions: Figure 5 shows the dip-positions of the interference dips as a function of gestrinone hapten and anti-gestrinone increasing concentrations. It was observed that for HRP-h-G concentrations above 5 µg/mL the interference dip shift remained unchanged, indicating that the biosensing surface was saturated and the signal was due solely to immobilization of the hapten-protein conjugate. The total dip wavenumber shift (Δw) after saturation was around 30 cm−1. A blocking step was necessary to deactivate the remaining epoxy groups on the surface before starting the antibody addition and, as expected, the blocking step with ethanolamine 0.1 M did not yield significant signal variations.


Biomolecular Interaction Analysis of Gestrinone-anti-Gestrinone Using Arrays of High Aspect Ratio SU-8 Nanopillars.

Ortega FJ, Bañuls MJ, Sanza FJ, Casquel R, Laguna MF, Holgado M, López-Romero D, Barrios CA, Maquieira Á, Puchades R - Biosensors (Basel) (2012)

Dip shift for HRP-h-G immobilization and anti-gestrinone antibody recognition for the dip centered at 15,300 cm−1. Each measurement is the average of five replicates.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4263551&req=5

biosensors-02-00291-f005: Dip shift for HRP-h-G immobilization and anti-gestrinone antibody recognition for the dip centered at 15,300 cm−1. Each measurement is the average of five replicates.
Mentions: Figure 5 shows the dip-positions of the interference dips as a function of gestrinone hapten and anti-gestrinone increasing concentrations. It was observed that for HRP-h-G concentrations above 5 µg/mL the interference dip shift remained unchanged, indicating that the biosensing surface was saturated and the signal was due solely to immobilization of the hapten-protein conjugate. The total dip wavenumber shift (Δw) after saturation was around 30 cm−1. A blocking step was necessary to deactivate the remaining epoxy groups on the surface before starting the antibody addition and, as expected, the blocking step with ethanolamine 0.1 M did not yield significant signal variations.

Bottom Line: After gestrinone antigen immobilization on the BICELLs, the immunorecognition was performed.The cells were interrogated vertically by using micron spot size Fourier transform visible and IR spectrometry (FT-VIS-IR), and the dip wavenumber shift was monitored.The biosensing assay exhibited good reproducibility and sensitivity (LOD = 0.75 ng/mL).

View Article: PubMed Central - PubMed

Affiliation: Centro de Reconocimiento Molecular y Desarrollo Tecnológico, Departamento de Química, Universitat Politècnica de València, Camino de Vera s/n, Valencia 46022, Spain. fraorhi1@upvnet.upv.es.

ABSTRACT
In this paper, label-free biosensing for antibody screening by periodic lattices of high-aspect ratio SU-8 nano-pillars (BICELLs) is presented. As a demonstration, the determination of anti-gestrinone antibodies from whole rabbit serum is carried out, and for the first time, the dissociation constant (KD = 6 nM) of antigen-antibody recognition process is calculated using this sensing system. After gestrinone antigen immobilization on the BICELLs, the immunorecognition was performed. The cells were interrogated vertically by using micron spot size Fourier transform visible and IR spectrometry (FT-VIS-IR), and the dip wavenumber shift was monitored. The biosensing assay exhibited good reproducibility and sensitivity (LOD = 0.75 ng/mL).

No MeSH data available.


Related in: MedlinePlus