High sensitive detection of C-reactive protein by total internal reflection fluorescence microscopy on rapidly making nanoarray protein chip

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Title
High sensitive detection of C-reactive protein by total internal reflection fluorescence microscopy on rapidly making nanoarray protein chip
Author(s)
M S Islam; Hee Gu Lee; J Choo; J M Song; S H Kang
Bibliographic Citation
Talanta, vol. 81, no. 4, pp. 1402-1408
Publication Year
2010
Abstract
This study investigated a method for the high sensitivity detection and quantification of the C-reactive protein (CRP) in human serum using total internal reflection fluorescence microscopy (TIRFM) on a rapidly made nanoarray protein chip. The nanoarray biotin-probe was patterned onto 3-mercaptopropyl trimethoxysilane-coated cover glass with a spot diameter of ∼400 nm within 1 min using a NanoeNabler™-based surface patterning tool. The unlabeled CRP molecules were detected in human sera using TIRFM, based on a sandwich fluorescence immunoassay. The linear regression for standard CRP in the range of 50 zM-1 fM was determined using the equation y = 0.437x + 84.991 (R = 0.9993). This proposed method was ∼2000 times faster than conventional atomic force microscopy based dip-pen nanolithography in terms of the chip manufacturing process. Additionally this method was 6 × 106 times more sensitive than enzyme-linked immunosorbent assay and exhibited a wide dynamic linear range (50 zM-1 fM).
Keyword
C-reactive proteinNanoarray protein chipSingle-molecule detectionSurface patterning toolTotal internal reflection fluorescence microscopy (TIRFM)
ISSN
0039-9140
Publisher
Elsevier
DOI
http://dx.doi.org/10.1016/j.talanta.2010.02.042
Type
Article
Appears in Collections:
Division of Biomedical Research > Immunotherapy Research Center > 1. Journal Articles
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