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Biosensors (Basel). 2015 Jan 19;5(1):27-36. doi: 10.3390/bios5010027.

Kinetic Analyses of Data from a Human Serum Albumin Assay Using the liSPR System.

Author information

1
Institute of Food Technology and Bioprocess Engineering, Technische Universität Dresden, Dresden 01062, Germany. anja.henseleit@tu-dresden.de.
2
Institute of Food Technology and Bioprocess Engineering, Technische Universität Dresden, Dresden 01062, Germany. carolin.pohl@tu-dresden.de.
3
QuoData GmbH, Prellerstraße 14, Dresden 01309, Germany. hans-michael.kaltenbach@quodata.de.
4
QuoData GmbH, Prellerstraße 14, Dresden 01309, Germany. hettwer@quodata.de.
5
New diagnostics GmbH, Moosstraße 92c, Freising D-85356, Germany. simon@new-diagnostics.com.
6
QuoData GmbH, Prellerstraße 14, Dresden 01309, Germany. uhlig@quodata.de.
7
Institute of Food Technology and Bioprocess Engineering, Technische Universität Dresden, Dresden 01062, Germany. nataliehaustein@yahoo.de.
8
Institute of Food Technology and Bioprocess Engineering, Technische Universität Dresden, Dresden 01062, Germany. thomas.bley@tu-dresden.de.
9
Institute of Food Technology and Bioprocess Engineering, Technische Universität Dresden, Dresden 01062, Germany. elke.boschke@tu-dresden.de.

Abstract

We used the interaction between human serum albumin (HSA) and a high-affinity antibody to evaluate binding affinity measurements by the bench-top liSPR system (capitalis technology GmbH). HSA was immobilized directly onto a carboxylated sensor layer, and the mechanism of interaction between the antibody and HSA was investigated. The bivalence and heterogeneity of the antibody caused a complex binding mechanism. Three different interaction models (1:1 binding, heterogeneous analyte, bivalent analyte) were compared, and the bivalent analyte model best fit the curves obtained from the assay. This model describes the interaction of a bivalent analyte with one or two ligands (A + L ↔ LA + L ↔ LLA). The apparent binding affinity for this model measured 37 pM for the first reaction step, and 20 pM for the second step.

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