Carbon nanofiber-based luminol-biotin probe for sensitive chemiluminescence detection of protein.
A carbon nanofiber-based luminol-biotin probe was synthesized for the sensitive chemiluminescence (CL) detection of a target protein by grafting luminol and biotin onto an oxidized carbon nanofiber. This carbon nanofiber was prepared by chemical vapor-deposition with methane in the presence of the Ni-Cu-MgO catalyst, which was followed by oxidization with HNO3-H2SO4 to produce a carboxyl group on the surface of the nanofiber. The material was grafted with luminol and biotin by means of a standard carbodiimide activation of COOH groups to produce corresponding amides. The substance was water-soluble and thus could be utilized as a sensitive CL probe for a protein assay. The probe showed highly specific affinity towards the biotin-labeled antibody via a streptavidin-biotin interaction. The detection limit for this model assay was approximately 0.2 pmol of the biotinized IgG spotted on a polyvinylidene fluoride (PVDF) membrane. Nonspecific binding to other proteins was not observed. Therefore, the synthesized carbon nanofiber-based CL probe may be useful for a sensitive and specific analysis of the target protein.
Department of Chemical Organic Technology and Petrochemistry, Faculty of Chemistry, Silesian University of Technology.
Stefan
S
Baj
Tomasz
T
Krawczyk
Natalia
N
Pradel
Md Golam
MG
Azam
Takayuki
T
Shibata
Shpend
S
Dragusha
Krzysztof
K
Skutil
Miroslawa
M
Pawlyta
Masaaki
M
Kai
D000818:Animals; D015374:Biosensing Techniques / Q000379:methods*; D001710:Biotin / Q000737:chemistry*; D002244:Carbon / Q000737:chemistry*; D006801:Humans; D007074:Immunoglobulin G / Q000032:analysis / Q000737:chemistry; D057230:Limit of Detection; D008163:Luminescent Measurements; D008165:Luminol / Q000737:chemistry*; D008567:Membranes, Artificial; D008697:Methane / Q000737:chemistry; D057139:Nanofibers / Q000737:chemistry*; D010084:Oxidation-Reduction; D011145:Polyvinyls / Q000737:chemistry; D011506:Proteins / Q000032:analysis* / Q000737:chemistry; D012995:Solubility; D014835:Volatilization; D005465:Fluorocarbon Polymers
D001710:Biotin; D002244:Carbon; D007074:Immunoglobulin G; D008165:Luminol; D008567:Membranes, Artificial; D008697:Methane; D011145:Polyvinyls; D011506:Proteins; C024865:polyvinylidene fluoride; D005465:Fluorocarbon Polymers