A novel method for monitoring monoclonal antibody production during cell culture.
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.
Szmacinski
Henryk
H
Center for Fluorescence Spectroscopy, Department of Biochemistry and Molecular Biology, University of Maryland Baltimore, 725 W. Lombard St., Baltimore, Maryland 21201, USA. henry@cfs.umbi.umd.edu
Smith
Derek S
DS
Hanson
Michael A
MA
Kostov
Yordan
Y
Lakowicz
Joseph R
JR
Rao
Govind
G
D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis* / Q000096:biosynthesis*; D018929:Cell Culture Techniques; D004797:Enzyme-Linked Immunosorbent Assay; D005453:Fluorescence; D015200:Fluoroimmunoassay; D005470:Fluorometry / Q000379:methods*; D006825:Hybridomas / Q000276:immunology; D007074:Immunoglobulin G / Q000032:analysis / Q000276:immunology; D007700:Kinetics; D051379:Mice
D000911:Antibodies, Monoclonal; D007074:Immunoglobulin G