Analysis of kinetic data of antibody-antigen interaction from an optical biosensor by exponential curve fitting.
An optical biosensor system employing a resonant mirror (RM), with a stirred cuvette has been used to follow the interaction of a recombinant antibody fragment with its antigen, hen egg lysozyme. The data generated by the biosensor were analysed in order to determine the kinetic constants for the interaction using a linear transform (derivative analysis). For comparison the data were also analysed using an exponential curve fitting routine. It was demonstrated that the exponential curve fitting method produced results which were in agreement with the existing linear transform method. It was also shown that early fitting of the association phase response, using the exponential curve fitting routine between 0 and 70 s after sample addition, yielded sufficient information to provide a prediction of Kon. The potential use of the optical biosensor for the rapid monitoring of protein production and purification is discussed.
Gill
A
A
Department of Chemical and Biochemical Engineering, University College London, UK.
Leatherbarrow
R J
RJ
Hoare
M
M
Pollard-Knight
D V
DV
Lowe
P A
PA
Fortune
D H
DH
D000818:Animals; D000906:Antibodies / Q000235:genetics; D000937:Antigen-Antibody Reactions / Q000502:physiology*; D015374:Biosensing Techniques; D001709:Biotechnology; D002645:Chickens; D003627:Data Interpretation, Statistical; D004800:Enzymes, Immobilized; D005260:Female; D007700:Kinetics; D009113:Muramidase / Q000276:immunology; D055095:Optics and Photonics / Q000295:instrumentation*; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology
D000906:Antibodies; D004800:Enzymes, Immobilized; D011994:Recombinant Proteins; D009113:Muramidase