Comparison of techniques for monitoring antibody fragment production in E. coli fermentation cultures.
The use of an optical biosensor for monitoring antibody fragment accumulation following induction in a batch fermentation of recombinant E. coli is compared to the more traditional method of ELISA quantification. Using the biosensor, concentration data can be obtained within minutes of sample addition to the device, compared to an average assay time of 3-4 h for the ELISA. We describe two biosensor assays developed as an alternative to ELISA and compare them with ELISA in the ability to provide quantitative product accumulation profiles during fermentation. Discrepancies in titers recorded by the assays are explained by a combination of differences in product variants detected by each assay and interference from sample contaminants. Method of sample preparation is also shown to be important if accurate concentration data is required. Both biosensor assays are shown to be capable of providing product accumulation profiles comparable to those obtained by ELISA. The use of a rapid extraction technique would allow such data to be obtained during process operation, enabling improved fermentation control and more rapid process development.
Bowering
Leigh C
LC
The Advanced Centre for Biochemical Engineering, Department of Biochemical Engineering, University College London, Torrington Place, London WC1E 7JE, UK.
Bracewell
Daniel G
DG
Kesharvarz-Moore
Eli
E
Hoare
Mike
M
Weir
A Neil C
AN
D000911:Antibodies, Monoclonal; D019149:Bioreactors; D015374:Biosensing Techniques / Q000379:methods* / Q000592:standards; D002138:Calibration; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000378:metabolism*; D005285:Fermentation; D006801:Humans; D007128:Immunoglobulin Fragments / Q000032:analysis* / Q000096:biosynthesis*; D013205:Staphylococcal Protein A
D000911:Antibodies, Monoclonal; D007128:Immunoglobulin Fragments; D013205:Staphylococcal Protein A