Novel hot electron-emitting working electrodes and conventional counter electrodes were created by screen printing. Thus, low-cost disposable electrode chips for bioaffinity assays were produced to replace our older expensive electrode chips manufactured by manufacturing techniques of electronics from silicon or on glass chips. The present chips were created by printing as follows: (i) silver lines provided the electronic contacts, counter electrode and the bottom of the working electrode and counter electrode, (ii) the composite layer was printed on appropriate parts of the silver layer, and (iii) finally a hydrophobic ring was added to produce the electrochemical cell boundaries. The applicability of these electrode chips in bioaffinity assays was demonstrated by an immunoassay of human C-reactive protein (i) using Tb(III) chelate label displaying long-lived hot electron-induced electrochemiluminescence (HECL) and (ii) now for the first time fluorescein isothiocyanate (FITC) was utilized as an a low-cost organic label displaying a short-lived HECL in a real-world bioaffinity assay.
A new label-free molecular probe for luminescent nucleotide detection in neutral aqueous solution is presented. Phosphate-containing molecules, such as nucleotides possess vital role in cell metabolism, energy economy, and various signaling processes. Thus, the monitoring of nucleotide concentration and nucleotide related enzymatic reactions is of high importance. Two component lanthanide complex formed from Tb(III) ion carrier and light harvesting antenna, readily distinguishes nucleotides containing different number of phosphates and enable direct detection of enzymatic reactions converting nucleotide triphosphate (NTP) to nucleotide di/monophosphate or the opposite. Developed sensor enables the detection of enzymatic activity with a low nanomolar sensitivity, as highlighted with K-Ras and apyrase enzymes in their hydrolysis assays performed in a high throughput screening compatible 384-well plate format.