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A novel method for monitoring monoclonal antibody production during cell culture
Author(s) -
Szmacinski Henryk,
Smith Derek S.,
Hanson Michael A.,
Kostov Yordan,
Lakowicz Joseph R.,
Rao Govind
Publication year - 2008
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.21795
Subject(s) - monoclonal antibody , bioprocess , biomolecule , substrate (aquarium) , chemistry , fluorescence , chromatography , analytical chemistry (journal) , biophysics , nanotechnology , biological system , antibody , materials science , chemical engineering , biochemistry , biology , optics , ecology , physics , engineering , immunology
Abstract We describe a new format for surface‐based fluoroimmunoassays that allows detection of biomolecule interactions without separation steps. The bioactive layer was immobilized on the surface of a glass substrate covered with silver islands that provide optical amplification of the distinctive fluorescence signal from bound probes when compared to unbound probes. The technique used was phase‐modulation fluorometry that allows sensitive detection of bound probes with a very short lifetime in the presence of excess free probes in solution. The new method was applied to assay monoclonal antibody production during cell culture. Excellent agreement was found between the new method and ELISA analysis of hybridoma cell culture samples. It is predicted that the near real time monitoring of protein products during bioprocessing will be possible with the described technology. Biotechnol. Bioeng. 2008;100: 448–457. © 2008 Wiley Periodicals, Inc.