Monitoring an antibody affinity chromatography with a label-free optical biosensor technique.
In the production of polyclonal antibody, a purification step is necessary which is often done by affinity chromatography. We present a biosensor system based on reflectometric interference spectroscopy (RIfS) to monitor the quantity and quality in terms of affinity and kinetic constants of the antibody during this procedure. Biosensors are rapid compared with ELISA, which is done in practice and can work fully automated. They provide additional information about the active antibody to protein concentration ratio and the affinity of the antibody. We show how to determine these values very accurate. In addition, we describe a new rapid method to monitor the affinity chromatography in process. This gives the possibility to select antibody fractions with best properties in respect to the application.
Proll
G
G
Institute of Physical and Theoretical Chemistry (IPTC), Eberhard-Karls-University of Tuebingen, Auf der Morgenstelle 8, D-72076 Tuebingen, Germany. Guenther.Proll@ipc.uni-tuebingen.de
Kumpf
M
M
Mehlmann
M
M
Tschmelak
J
J
Griffith
H
H
Abuknesha
R
R
Gauglitz
G
G
D000915:Antibody Affinity; D015374:Biosensing Techniques / Q000379:methods*; D002846:Chromatography, Affinity; D004797:Enzyme-Linked Immunosorbent Assay; D013057:Spectrum Analysis