Publication details

Nanopolyperoxidase – a universal system for simultaneous colorimetric signal enhancement and nanoscale visualization of antibody labeling



Year of publication 2008
Type Article in Proceedings
Conference 5th international interdisciplinary meeting on bioanalysis CECE 2008
MU Faculty or unit

Faculty of Science

Field Biochemistry
Keywords antibody enzyme-label blotting ELISA
Description During past decades, a variety of signal amplification approaches in heterogeneous affinity immunoassays (including ELISA) has been developed. Those procedures are usually based either on recirculation enzymatic or repeated multiplication of an enzyme label via avidin-biotin system. From practical point of view, the multiplication procedure should not change experimental part of method, e.g. number of steps in procedure or solution content should not be changed. For this reason, an idea of nanopolyperoxidase (NPP) has originated. The principle is simple: gold nanoparticles are modified with an antibody as well as with labeling enzyme (Horseradish peroxidase; ration 2:8). Modified nanoparticle is stabilized in a PEG solution. Prepared NPP was studied using various methods. Atomic Force Microscopy (AFM) was used for characterization of structure and morphology in a nanometer scale (see image below). Use of NPP in construction of competitive ELISA as well as in visualization of a protein (Human Serum Albumin as model compound) on a blotting membrane is presented.
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