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

Biosensors

Anja Henseleit³  Carolin Pohl³  Hans-Michael Kaltenbach²  Karina Hettwer²  Kirsten Simon¹  Steffen Uhlig²  Natalie Haustein³  Thomas Bley³  Elke Boschke³ 

[1] New diagnostics GmbH, Moosstraße 92c, Freising D-85356, Germany; E-Mail:;QuoData GmbH, Prellerstraße 14, Dresden 01309, Germany; E-Mails:;Institute of Food Technology and Bioprocess Engineering, Technische Universität Dresden, Dresden 01062, Germany; E-Mails:

关键词: surface plasmon resonance (SPR);    human serum albumin (HSA);    antibody;    bivalent analyte;   

DOI  :  10.3390/bios5010027

来源: mdpi

【 摘 要 】

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.

【 授权许可】

CC BY   
© 2015 by the authors; licensee MDPI, Basel, Switzerland.