4.8 Article

Extended Nanofluidic Immunochemical Reaction with Femtoliter Sample Volumes

Journal

SMALL
Volume 10, Issue 8, Pages 1514-1522

Publisher

WILEY-V C H VERLAG GMBH
DOI: 10.1002/smll.201302709

Keywords

extended nanospace; immunoassay; surface patterning; nanofluidics

Funding

  1. [23-9195]
  2. Grants-in-Aid for Scientific Research [11J09195] Funding Source: KAKEN

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The growing need to optimize immunoassay performance driven by interest in analyzing individual cells has resulted in a decrease in the amount of sample required. Miniaturized immunoassays that use ultra-small femtoliter to attoliter sample volumes, a range known as the extended nanospace, can satisfy this analytical need; however, capturing every targeted molecule without loss in extended nanochannels for subsequent detection remains challenging. This is the first report of a successful extended nanofluidics-based quantitative immunochemical reaction capable of high capture efficiency using a femtoliter-scale sample volume. A novel patterning method using a photolithographic technique with vacuum ultraviolet light and low-temperature (100 degrees C) bonding enables patterning of functional groups for antibody immobilization before bonding, resulting in an immunochemical reaction space of only 86 fL. Reaction rate analyses indicate a decrease in the required sample volume to 810 fL and improvement in the limit of detection to 3 zmol, 5-6 orders of magnitude better than possible with the microfluidic immunoassay format. Highly efficient (near 100%) immunochemical reactions on a seconds time scale are possible due to the nm-scale diffusion length, which should be advantageous for the analysis of ultra-low-volume samples.

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