TY  - JOUR
A1  - Cywinski, Piotr J.
A1  - Pietraszkiewicz, Marek
A1  - Maciejczyk, Michal
A1  - Gorski, Krzysztof
A1  - Hammann, Tommy
A1  - Liermann, Konstanze
A1  - Paulke, Bernd-Reiner
A1  - Löhmannsröben, Hans-Gerd
T1  - Total protein concentration quantification using nanobeads with a new highly luminescent terbium(III) complex
JF  - RSC Advances
N2  - Total protein concentration (TPC) is a key parameter in many biochemical experiments and its quantification is often necessary for isolation, separation, and analysis of proteins. A sensitive and rapid nanobead-based TPC quantification assay based on Forster Resonance Energy Transfer (FRET) has been developed. A new, highly luminescent Tb(III) complex has been synthesised and applied as donor in this FRET assay with an organic dye (Cy5) as acceptor. FRET-induced changes in luminescence have been investigated both at donor and acceptor emission wavelength using time-resolved luminescence spectroscopy with time-gated detection. In the assay, the Tb(III) complex and fine-tuned polyglycidyl methacrylate (PGMA) nanobeads ensure that an improvement in sensitivity and background reduction is achieved. Using 40 nm large PGMA nanobeads loaded with the Tb(III) complex, it is possible to determine TPC down to 50 ng mL(-1) in just 10 minutes. Through specific assay components the sensitivity has been improved when compared to existing nanobead-based assays and to currently known commercial methods. Additionally, the assay is relatively insensitive to the presence of contaminants, such as non-ionic detergents commonly found in biological samples. Due to no need for any centrifugal steps, this mix-and-measure bioassay can easily be implemented into routine TPC quantification protocols in biochemical laboratories.
Y1  - 2016
U6  - https://doi.org/10.1039/c6ra23207h
SN  - 2046-2069
VL  - 6
SP  - 115068
EP  - 115073
PB  - Royal Society of Chemistry
CY  - Cambridge
ER  -