title: Kinetic Analyses of Data from a Human Serum Albumin Assay Using the liSPR System.
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.
authors.affiliation: Institute of Food Technology and Bioprocess Engineering, Technische Universität Dresden, Dresden 01062, Germany. anja.henseleit@tu-dresden.de.
authors.forename: Anja
authors.initials: A
authors.lastname: Henseleit
authors.affiliation: Institute of Food Technology and Bioprocess Engineering, Technische Universität Dresden, Dresden 01062, Germany. carolin.pohl@tu-dresden.de.
authors.forename: Carolin
authors.initials: C
authors.lastname: Pohl
authors.affiliation: QuoData GmbH, Prellerstraße 14, Dresden 01309, Germany. hans-michael.kaltenbach@quodata.de.
authors.forename: Hans-Michael
authors.initials: HM
authors.lastname: Kaltenbach
authors.affiliation: QuoData GmbH, Prellerstraße 14, Dresden 01309, Germany. hettwer@quodata.de.
authors.forename: Karina
authors.initials: K
authors.lastname: Hettwer
authors.affiliation: New diagnostics GmbH, Moosstraße 92c, Freising D-85356, Germany. simon@new-diagnostics.com.
authors.forename: Kirsten
authors.initials: K
authors.lastname: Simon
authors.affiliation: QuoData GmbH, Prellerstraße 14, Dresden 01309, Germany. uhlig@quodata.de.
authors.forename: Steffen
authors.initials: S
authors.lastname: Uhlig
authors.affiliation: Institute of Food Technology and Bioprocess Engineering, Technische Universität Dresden, Dresden 01062, Germany. nataliehaustein@yahoo.de.
authors.forename: Natalie
authors.initials: N
authors.lastname: Haustein
authors.affiliation: Institute of Food Technology and Bioprocess Engineering, Technische Universität Dresden, Dresden 01062, Germany. thomas.bley@tu-dresden.de.
authors.forename: Thomas
authors.initials: T
authors.lastname: Bley
authors.affiliation: Institute of Food Technology and Bioprocess Engineering, Technische Universität Dresden, Dresden 01062, Germany. elke.boschke@tu-dresden.de.
authors.forename: Elke
authors.initials: E
authors.lastname: Boschke