albumin - publications

Predict more albumin - ligand interactions now!

1. Talanta. 2012 May 30;94:65-9. Epub 2012 Mar 1.

A rational route to the development of a competitive capillary electrophoresis
immunoassay: Assessment of the variables affecting the performances of a
competitive capillary electrophoresis immunoassay for human serum albumin.

Giovannoli C, Baggiani C, Passini C, Biagioli F, Anfossi L, Giraudi G.

Laboratory of Bioanalytical Chemistry, Department of Chemistry, University of
Torino, via Giuria 5, 10125 Torino, Italy.

Affinity capillary electrophoresis is a powerful analytical tool to extract
quantitative information about the binding properties of different interacting
systems. The use of LIF detection makes the technique suitable for screening
strong binding interactions. The non-equilibrium electrophoretic separations of
pre-equilibrated mixtures of ligand and receptor are generally used for such
strong molecular interactions allowing the assessment of capillary
electrophoresis immunoassays, mostly in competitive formats. As the analytical
performances of the assay strongly depend on the preservation of the binding
properties during the separation, a rational route to assay development has to be
followed to get the best conditions. The paper describes the steps followed to
set-up a competitive immunoassay for human serum albumin (HSA) by using a labeled
protein (HSA-FITC) and an anti-HSA polyclonal antiserum. A labeling degree of
around 1 of the HSA-FITC conjugates is needed to get narrow electrophoretic peak
while the titration curve is used to define the optimal antiserum dilution. An
antiserum-labeled protein affinity constant of 1.34×10(7)M(-1) was measures in
the selected separation conditions. Furthermore, in order to maximize the assay
competition between the labeled and unlabelled HSA a short pre-incubation step of
the antiserum with the unlabelled HSA (the analyte) was introduced to promote a
sharp increase in assay sensitivity.

Copyright © 2012 Elsevier B.V. All rights reserved.

PMID: 22608415 [PubMed - in process]