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Factorization of preparative protein chromatograms with hard-constraint multivariate curve resolution and second-derivative pretreatment

Rüdt, Matthias; Andris, Sebastian; Schiemer, Robin; Hubbuch, Jürgen

Abstract:
Current biopharmaceutical production heavily relies on chromatography for protein purification. Recently, research has intensified towards finding suitable solutions to monitoring the chromatographic steps by multivariate spectroscopic sensors. Here, hard-constraint multivariate curve resolution (MCR) was investigated as a calibration-free method for factorizing bilinear preparative protein chromatograms into concentrations and spectra. Protein elutions were assumed to follow exponentially modified Gaussian (EMG) curves. In three case studies, MCR was applied to chromatograms of second-derivative ultraviolet and visible (UV–vis) spectra. The three case studies consisted of the separation of a ternary mixture (ribonuclease A, cytochrome c, and lysozyme), multiple binary chromatography runs of cytochrome c and lysozyme, and the separation of an antibody–drug conjugate (ADC) from unconjugated immunoglobulin G (IgG). In all case studies, good estimates of the elution curves were obtained. R2 values compared to off-line analytics exceeded 0.90. The estimated spectra allowed for protein identification based on a protein spectral library. In summary, MCR was shown to be well able to factorize protein chromatograms without prior calibration. ... mehr


Verlagsausgabe §
DOI: 10.5445/IR/1000091037
Veröffentlicht am 14.02.2019
Originalveröffentlichung
DOI: 10.1016/j.chroma.2018.11.065
Scopus
Zitationen: 2
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Zitationen: 2
Cover der Publikation
Zugehörige Institution(en) am KIT Institut für Bio- und Lebensmitteltechnik (BLT)
Publikationstyp Zeitschriftenaufsatz
Publikationsmonat/-jahr 01.2019
Sprache Englisch
Identifikator ISSN: 0021-9673, 0376-737x, 0378-4355
urn:nbn:de:swb:90-910371
KITopen-ID: 1000091037
Erschienen in Journal of chromatography
Verlag Elsevier
Band 1585
Seiten 152–160
Projektinformation nextBioPharmDSP (EU, H2020, 635557)
Nachgewiesen in Web of Science
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Scopus
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