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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 against antibody concentration for bovine serum albumin (BSA)-anti-BSA and gestrinone-anti-gestrinone systems.
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biosensors-02-00291-f006: Dip shift against antibody concentration for bovine serum albumin (BSA)-anti-BSA and gestrinone-anti-gestrinone systems.

Mentions: Figure 6 shows the different assay responses (BSA/ anti-BSA and gestrinone/anti-gestrinone) using the same optical transduction (SU-8 nanopillars over one micron of SiO2 and Si substrate). It can be seen how, when the sensing surface is saturated with antibodies, the signal level reaches a similar wavenumber displacement. In the case of anti-gestrinone, the slope was higher than for anti-BSA, and thus, the antibody concentration where 50% of the species are associated (value of KD) resulted higher for anti-BSA (KD = 33 nM) than for anti-gestrinone (KD = 6 nM). Then, the affinity was calculated as the inverse of KD resulting in 0.03 nM−1 and 0.17 nM−1 for anti-BSA and anti-gestrinone, respectively. These values agree with the data reported in the literature for these systems [23,35,36,37] and demonstrate the applicability of the sensor for the determination of binding constants or biorecognition extension.


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 against antibody concentration for bovine serum albumin (BSA)-anti-BSA and gestrinone-anti-gestrinone systems.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

biosensors-02-00291-f006: Dip shift against antibody concentration for bovine serum albumin (BSA)-anti-BSA and gestrinone-anti-gestrinone systems.
Mentions: Figure 6 shows the different assay responses (BSA/ anti-BSA and gestrinone/anti-gestrinone) using the same optical transduction (SU-8 nanopillars over one micron of SiO2 and Si substrate). It can be seen how, when the sensing surface is saturated with antibodies, the signal level reaches a similar wavenumber displacement. In the case of anti-gestrinone, the slope was higher than for anti-BSA, and thus, the antibody concentration where 50% of the species are associated (value of KD) resulted higher for anti-BSA (KD = 33 nM) than for anti-gestrinone (KD = 6 nM). Then, the affinity was calculated as the inverse of KD resulting in 0.03 nM−1 and 0.17 nM−1 for anti-BSA and anti-gestrinone, respectively. These values agree with the data reported in the literature for these systems [23,35,36,37] and demonstrate the applicability of the sensor for the determination of binding constants or biorecognition extension.

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