Real time optical immunosensing with flow-through porous alumina membranes

Jesús Álvarez; Laura Sola; Marina Cretich; Marcus J. Swann; Kristinn B. Gylfasson; Tormod Volden; Marcella Chiari; Daniel Hill

doi:10.1016/j.snb.2014.06.027

Real time optical immunosensing with flow-through porous alumina membranes

Jesús Álvarez
, Laura Sola
, Marina Cretich
, Marcus J. Swann
, Kristinn B. Gylfasson
, Tormod Volden
, Marcella Chiari
, Daniel Hill^*

^*Corresponding author for this work

Aston Institute of Photonic Technologies (AiPT)

Universitat de València
Consiglio Nazionale delle Ricerche
Farfield Group Ltd.
Kungliga Tekniska Högskolan
Central Switzerland Center

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Abstract

Through the presentation of analytical data from bioassay experiments, measured by polarimetry, we demonstrate for the first time a real time immunoassay within a free standing macroporous alumina membrane. The 200 nm nominal pore diameter of the membrane enables flow-through, thereby providing an ideal fluidic platform for the targeted delivery of analytes to bioreceptors immobilized on the pore walls, enabling fast sensing response times and the use of small sample volumes (<100 μL). For the immunoassay, the pore walls were first coated with the functional copolymer, copoly(DMA-NAS) using a novel coupling process, before immobilization of the allergen protein, β-lactoglobulin, by spotting. The immuno-assay then proceeded with the binding of the primary and secondary antibody cognates, rabbit anti-β-lactoglobulin and anti-rabbit IgG respectively. Through the use of streptavidin coated quantum dots as refractive index signal enhancers, a noise floor for individual measurements of 3.7 ng/mL (25 pM) was obtained, with an overall statistical, or formal assay LOD of 33.7 ng/mL (225 pM), for total assay time below 1 h.

Original language	English
Pages (from-to)	834-839
Number of pages	6
Journal	Sensors and Actuators, B: Chemical
Volume	202
DOIs	https://doi.org/10.1016/j.snb.2014.06.027
Publication status	Published - 31 Oct 2014

Bibliographical note

Funding

The authors would like to acknowledge the contributions from Geoff Platt (Farfield), Isabelle Metton (Phylogene), Steven Sievers (Charité) and Thomas Stadelmann (CSEM) for their participating in prior biosensing experiments that led up to this work. We also thank Helmut Knapp, Siegfried Graf (both CSEM) for the design and fabrication of the flow cell and Wouter van der Wijngaart (KTH) for fluidic discussions. Finally we thank Intenanomat for the use of their wet labs for reagent preparation and storage. This work was carried out within the FP7-ICT-257401-POSITIVE project, funded by the European Commission .

Keywords

Copolymer
Form birefringence
Optical biosensing
Polarimetry
Porous alumina
Quantum dots

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10.1016/j.snb.2014.06.027

Real time optical immunosensing with flow-through porous alumina membranes
© 2014, Elsevier. Licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International http://creativecommons.org/licenses/by-nc-nd/4.0/
Accepted author manuscript, 2.04 MBLicence: CC BY-NC-ND 3.0

Cite this

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title = "Real time optical immunosensing with flow-through porous alumina membranes",

abstract = "Through the presentation of analytical data from bioassay experiments, measured by polarimetry, we demonstrate for the first time a real time immunoassay within a free standing macroporous alumina membrane. The 200 nm nominal pore diameter of the membrane enables flow-through, thereby providing an ideal fluidic platform for the targeted delivery of analytes to bioreceptors immobilized on the pore walls, enabling fast sensing response times and the use of small sample volumes (<100 μL). For the immunoassay, the pore walls were first coated with the functional copolymer, copoly(DMA-NAS) using a novel coupling process, before immobilization of the allergen protein, β-lactoglobulin, by spotting. The immuno-assay then proceeded with the binding of the primary and secondary antibody cognates, rabbit anti-β-lactoglobulin and anti-rabbit IgG respectively. Through the use of streptavidin coated quantum dots as refractive index signal enhancers, a noise floor for individual measurements of 3.7 ng/mL (25 pM) was obtained, with an overall statistical, or formal assay LOD of 33.7 ng/mL (225 pM), for total assay time below 1 h.",

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AU - Álvarez, Jesús

AU - Sola, Laura

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AU - Swann, Marcus J.

AU - Gylfasson, Kristinn B.

AU - Volden, Tormod

AU - Chiari, Marcella

AU - Hill, Daniel

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N2 - Through the presentation of analytical data from bioassay experiments, measured by polarimetry, we demonstrate for the first time a real time immunoassay within a free standing macroporous alumina membrane. The 200 nm nominal pore diameter of the membrane enables flow-through, thereby providing an ideal fluidic platform for the targeted delivery of analytes to bioreceptors immobilized on the pore walls, enabling fast sensing response times and the use of small sample volumes (<100 μL). For the immunoassay, the pore walls were first coated with the functional copolymer, copoly(DMA-NAS) using a novel coupling process, before immobilization of the allergen protein, β-lactoglobulin, by spotting. The immuno-assay then proceeded with the binding of the primary and secondary antibody cognates, rabbit anti-β-lactoglobulin and anti-rabbit IgG respectively. Through the use of streptavidin coated quantum dots as refractive index signal enhancers, a noise floor for individual measurements of 3.7 ng/mL (25 pM) was obtained, with an overall statistical, or formal assay LOD of 33.7 ng/mL (225 pM), for total assay time below 1 h.

AB - Through the presentation of analytical data from bioassay experiments, measured by polarimetry, we demonstrate for the first time a real time immunoassay within a free standing macroporous alumina membrane. The 200 nm nominal pore diameter of the membrane enables flow-through, thereby providing an ideal fluidic platform for the targeted delivery of analytes to bioreceptors immobilized on the pore walls, enabling fast sensing response times and the use of small sample volumes (<100 μL). For the immunoassay, the pore walls were first coated with the functional copolymer, copoly(DMA-NAS) using a novel coupling process, before immobilization of the allergen protein, β-lactoglobulin, by spotting. The immuno-assay then proceeded with the binding of the primary and secondary antibody cognates, rabbit anti-β-lactoglobulin and anti-rabbit IgG respectively. Through the use of streptavidin coated quantum dots as refractive index signal enhancers, a noise floor for individual measurements of 3.7 ng/mL (25 pM) was obtained, with an overall statistical, or formal assay LOD of 33.7 ng/mL (225 pM), for total assay time below 1 h.

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Real time optical immunosensing with flow-through porous alumina membranes

Abstract

Bibliographical note

Funding

Keywords

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