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Rapid protein expression analysis with an interferometric biosensor for monitoring protein production

 
: Hoffmann, C.; Schmitt, K.; Brandenburg, A.; Hartmann, S.

:

Analytical and bioanalytical chemistry 387 (2007), Nr.5, S.1921-1932
ISSN: 1618-2642 (Print)
ISSN: 1618-2650 (Online)
Englisch
Zeitschriftenaufsatz
Fraunhofer IPM ()
surface plasmon resonance; self-assembled monolayer; label-free; molecular recognition; optical biosensor; grating coupler; sensor

Abstract
The concentration of a recombinantly expressed protein has to be monitored to select optimal expression conditions throughout the protein production process. Today this is usually achieved semiquantitatively with sodium dodecyl sulfate polyacrylamide gel electrophoresis/western blotting or with ELISAs, which are time- and labor-intensive methods. In this paper the applicability of a label-free sensor system based on a Young interferometer is presented as an alternative for the monitoring of recombinant protein production. Once a protein is successfully produced, the interferometric biosensor allows any protein-protein interaction to be characterized in a label-free manner. This is demonstrated with an antibody/antigen pair, where the antibody is directed against a four-amino-acid tag used for protein expression analysis as well as purification during recombinant prote in production. Label-free detection of the tagged protein is shown both in buffer and in bacterial cell lysate as a sample matrix. The system exhibiting a low limit of detection, low drift and reliable operation is compared with a commercial surface plasmon resonance sensor and a competitive ELISA.

: http://publica.fraunhofer.de/dokumente/N-60089.html