In situ NIR spectroscopy monitoring of plasmid production processes: effect of producing strain, medium composition and the cultivation strategy. Issue 2 (17th June 2014)

• Record Type: Journal Article
• Authors: Lopes, Marta B.; Gonçalves, Geisa A. L.; Felício‐Silva, Daniel; Prather, Kristala L. J.; Monteiro, Gabriel A.; Prazeres, Duarte M. F.; Calado, Cecília R. C.
• Is Part Of: Journal of chemical technology & biotechnology. Volume 90:Issue 2(2015:Feb.)
• Page Start: 255
• Page End: 261
• Publication Date: 2014-06-17
• Subjects: Biotechnology -- Periodicals; Chemistry, Technical -- Periodicals; Chemical engineering -- Periodicals; Industries -- Environmental aspects -- Periodicals; 660
• DOI: 10.1002/jctb.4431
• Languages: English
• ISSNs: 0268-2575

Abstract

<abstract abstract-type="main" id="jctb4431-abs-0001"> <title>Abstract</title> <sec id="jctb4431-sec-0001" sec-type="section"> <title>BACKGROUND</title> <p id="jctb4431-para-0001">While the pharmaceutical industry keeps an eye on plasmid DNA production for new generation gene therapies, real‐time monitoring techniques for plasmid bioproduction are as yet unavailable. This work shows the possibility of <italic>in situ</italic> monitoring of plasmid production in <italic>Escherichia coli</italic> cultures using a near infrared (NIR) fiber optic probe.</p> </sec> <sec id="jctb4431-sec-0002" sec-type="section"> <title>RESULTS</title> <p id="jctb4431-para-0002">Partial least squares (PLS) regression models based on the NIR spectra were developed for predicting bioprocess critical variables such as the concentrations of biomass, plasmid, carbon sources (glucose and glycerol) and acetate. In order to achieve robust models able to predict the performance of plasmid production processes, independently of the composition of the cultivation medium, cultivation strategy (batch versus fed‐batch) and <italic>E. coli</italic> strain used, three strategies were adopted, using: (i) <italic>E. coli</italic> DH5α cultures conducted under different media compositions and culture strategies (batch and fed‐batch); (ii) engineered <italic>E. coli</italic> strains, MG1655<italic>ΔendA</italic><italic>ΔrecAΔpgi</italic> and MG1655<italic>ΔendAΔrecA</italic>, grown on the same medium and culture strategy; (iii) diverse <italic>E. coli</italic> strains, over batch and fed‐batch cultivations and using different media compositions. PLS models showed high accuracy for predicting all variables in the three groups of cultures.</p> </sec> <sec id="jctb4431-sec-0003" sec-type="section"> <title>CONCLUSION</title> <p id="jctb4431-para-0003">NIR spectroscopy combined with PLS modeling provides a fast, inexpensive and contamination‐free technique to accurately monitoring plasmid bioprocesses in real time, independently of the medium composition, cultivation strategy and the <italic>E. coli</italic> strain used. © 2014 Society of Chemical Industry</p> </sec> </abstract>