High sensitive detection of C-reactive protein by total internal reflection fluorescence microscopy on rapidly making nanoarray protein chip.
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 approximately 400nm within 1min 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 50zM-1fM was determined using the equation y=0.437x+84.991 (R=0.9993). This proposed method was approximately 2000 times faster than conventional atomic force microscopy based dip-pen nanolithography in terms of the chip manufacturing process. Additionally this method was 6 x 10(6) times more sensitive than enzyme-linked immunosorbent assay and exhibited a wide dynamic linear range (50zM-1fM).
Islam
Md Shahinul
MS
Department of Chemistry and Research Institute of Physics and Chemistry (RINPAC), Chonbuk National University, Jeonju, South Korea.
Lee
Hee Gu
HG
Choo
Jaebum
J
Song
Joon Myong
JM
Kang
Seong Ho
SH
D001710:Biotin / Q000737:chemistry; D002097:C-Reactive Protein / Q000737:chemistry*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D004867:Equipment Design; D006801:Humans; D018625:Microscopy, Atomic Force / Q000379:methods; D008856:Microscopy, Fluorescence / Q000379:methods*; D036103:Nanotechnology / Q000379:methods*; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D017646:Organosilicon Compounds; D015203:Reproducibility of Results; D012821:Silanes / Q000737:chemistry; D013499:Surface Properties
D017646:Organosilicon Compounds; D012821:Silanes; D001710:Biotin; C102833:(3-mercaptopropyl)trimethoxysilane; D002097:C-Reactive Protein