Atomic absorption spectrometric detection of biotin-cymantrene as a metallo-tracer for the avidin–biotin system

Abstract

Biotin-cyclopentadienylmanganese tricarbonyl (biotin-cymantrene; biotin-Cy) is proposed as a universal metallo-tracer for immunoassays. Associated with the possibility of detecting this type of label (organometallic moieties) along with different analytical procedures, this reagent appears to be a universal system with the advantages of being inexpensive, stable and directly detected (no substrate is required). The optimum conditions for detecting the organometallic label biotin-Cy with a Zeeman atomic absorption spectrometer and the affinity of biotin-Cy for the streptavidin ligand immobilized on the walls of microtitre plates, wells or tubes are described. The results obtained demonstrate the possibility of assaying a wide range of clinical parameters because the detection limit of biotin-Cy is about 1 pg per assay. Based on the competition between immobilized bovine serum albumin adsorbed on the wall and free bovine serum albumin for specific polyclonal antibodies, the streptavidin-metallo-biotin system was tested on a competitive immunoassay model and a satisfactory calibration graph was obtained.

Ferrocenylethyl Phosphate: An Improved Substrate for the Detection of Alkaline Phosphatase by Cathodic Stripping Ion-Exchange Voltammetry. Application to the Electrochemical Enzyme Affinity Assay of Avidin

“…Heterogeneous Enzyme Assay of Avidin. The development of avidin−biotin technology has unified and simplified a variety of affinity sensing methods, and the avidin−biotin couple has been introduced in various indirect labeling procedures for immunoassays. ,, The first reason is the remarkable specific affinity between the two molecules (dissociation constant of ∼10 -15 ) . A second reason is the presence of four identical subunits on avidin, each of them being able to bind one biotin, and so an amplified signal can be generated.…”

Ultrafast 2DIR probe of a host-guest inclusion complex: Structural and dynamical constraints of nanoconfinement

“…CMT and its derivatives are stable, finding use as an anti-knocking additive in gasoline. 22 CMT has been investigated in the form of bioconjugates with biotin [23][24][25] and with cell penetrating peptides as a means of introducing cytotoxic carbon monoxide to cancer cells. We are investigating the suitability of a suite of metal carbonyl complexes for use as biophysical probes, including cymantrene due to its relatively wide array of proven applications.…”