Measurement and monitoring of the production of proteins during cell culture using plasmonic structures

A new method for surface-based fluoroimmunoassays that eliminates separation steps while still allowing high sensitivity detection of biomolecular interactions is presented. The capture antibody is electrostatically immobilized on a glass slide coated with a high density silver island film. The metal-enhanced fluorescence generated by the presence of the islands allows the sensitive detection of bound reporter antibodies versus those free in solution. In order to perform the measurement, phase-modulation fluorometry is employed which allows observation of the distinct fluorescence signal of the bound antibodies with a shorter lifetime than unbound antibodies. Here, we show the use of metal-enhanced fluorescence with phase-modulation fluorometry to quantify monoclonal antibody from a cell culture. The results show the new technique produces very similar data upon analysis as measured with ELISA analysis. With further optimization of the procedures, it is forecast that real time monitoring during bioprocessing will be feasible with the described technique.

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