Development of a single-step immunoassay microdevice based on a graphene oxide-containing hydrogel possessing fluorescence quenching and size separation functions

An immunoassay, which is an indispensable analytical method both in biological research and in medical fields was successfully integrated into a "single-step" by developing a microdevice composed of a graphene oxide (GO)-containing hydrogel and a poly (dimethylsiloxane) (PDMS) microchannel...

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Bibliographic Details
Published in:Analyst (London) 2017-01, Vol.142 (3), p.472-477
Main Authors: Shirai, Akihiro, Nakashima, Kaho, Sueyoshi, Kenji, Endo, Tatsuro, Hisamoto, Hideaki
Format: Article
Language:English
Subjects:
Antibodies
Antibodies - chemistry
Fluorescence
Fluorescence Resonance Energy Transfer
Graphene
Graphite
Humans
Hydrogels
Immunoassay
Immunoglobulin M - blood
Oxides
Polyethylene glycol
Polyethylene Glycols
Silicone resins
Surface chemistry
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Summary:An immunoassay, which is an indispensable analytical method both in biological research and in medical fields was successfully integrated into a "single-step" by developing a microdevice composed of a graphene oxide (GO)-containing hydrogel and a poly (dimethylsiloxane) (PDMS) microchannel array with a polyethylene glycol (PEG) coating containing a fluorescently-labelled antibody. Here we used 2-hydroxyethylmethacrylate (HEMA) as a monomer that is easily, and homogeneously, mixed with GO to synthesize the hydrogel. The fluorescence quenching and size separation functions were then optimized by controlling the ratios of HEMA and GO. Free fluorescently-labelled antibody was successfully separated from the immunoreaction mixture by the hydrogel network structure, and the fluorescence was subsequently quenched by GO. In comparison to the previously reported immunoassay system using GO, the present system achieved a very high fluorescence resonance energy transfer (FRET) efficiency (∼90%), due to the use of direct adsorption of the fluorescently-labelled antibody to the GO surface; in contrast, the former reported method relied on indirect adsorption of the fluorescently-labelled antibody via immunocomplex formation at the GO surface. Finally, the single-step immunoassay microdevice was made by combining the developed hydrogel and the PDMS microchannel with a coating containing the fluorescently-labelled antibody, and successfully applied for the single-step analysis of IgM levels in diluted human serum by simple introduction of the sample via capillary action.
ISSN:0003-2654
1364-5528
DOI:10.1039/c6an02485h