KIT | KIT-Bibliothek | Impressum | Datenschutz

Real-time monitoring and control of the load phase of a protein A capture step

Rüdt, Matthias 1; Brestrich, Nina 1; Rolinger, Laura 1; Hubbuch, Jürgen ORCID iD icon 1
1 Karlsruher Institut für Technologie (KIT)

Abstract:

The load phase in preparative Protein A capture steps is commonly not controlled in real‐time. The load volume is generally based on an offline quantification of the monoclonal antibody (mAb) prior to loading and on a conservative column capacity determined by resin‐life time studies. While this results in a reduced productivity in batch mode, the bottleneck of suitable real‐time analytics has to be overcome in order to enable continuous mAb purification. In this study, Partial Least Squares Regression (PLS) modeling on UV/Vis absorption spectra was applied to quantify mAb in the effluent of a Protein A capture step during the load phase. A PLS model based on several breakthrough curves with variable mAb titers in the HCCF was successfully calibrated. The PLS model predicted the mAb concentrations in the effluent of a validation experiment with a root mean square error (RMSE) of 0.06 mg/mL. The information was applied to automatically terminate the load phase, when a product breakthrough of 1.5 mg/mL was reached. In a second part of the study, the sensitivity of the method was further increased by only considering small mAb concentrations in the calibration and by subtracting an impurity background signal. ... mehr


Verlagsausgabe §
DOI: 10.5445/IR/1000060325
Veröffentlicht am 20.04.2018
Originalveröffentlichung
DOI: 10.1002/bit.26078
Scopus
Zitationen: 46
Web of Science
Zitationen: 41
Dimensions
Zitationen: 47
Cover der Publikation
Zugehörige Institution(en) am KIT Institut für Bio- und Lebensmitteltechnik (BLT)
Publikationstyp Zeitschriftenaufsatz
Publikationsmonat/-jahr 02.2017
Sprache Englisch
Identifikator ISSN: 0006-3592, 0368-1467, 1097-0290
urn:nbn:de:swb:90-603254
KITopen-ID: 1000060325
Erschienen in Biotechnology & bioengineering
Verlag John Wiley and Sons
Band 114
Heft 2
Seiten 368-373
Projektinformation nextBioPharmDSP (EU, H2020, 635557)
Vorab online veröffentlicht am 21.09.2016
Nachgewiesen in Dimensions
Scopus
Web of Science
KIT – Die Forschungsuniversität in der Helmholtz-Gemeinschaft
KITopen Landing Page