Carbon Nanofiber-based Luminol-biotin Probe for Sensitive Chemiluminescence Detection of Protein

Abstract

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.

Hexamethylene‐1,3‐bis(cetyldimethylammonium bromide) protected low‐toxicity ZnSe quantum dots as fluorescent probe for proteins

“…Many methods, such as resonance light scattering techniques [5][6][7], chemiluminescence [8], Bradford assay method [9], circular dichroism, fluorescence spectroscopy [10], Fourier transform infrared spectroscopy [11] and differential scanning calorimetry [12], have been proposed for the detection of proteins. Expensive instruments or complicated sample preparation processes are required, although these methods are sensitive.…”