Titlebar

Bibliografische Daten exportieren
Literatur vom gleichen Autor
plus auf ERef Bayreuth
plus bei Google Scholar

 

Hyphenation of multi-dimensional chromatography and mass spectrometry for the at-line-analysis of the integrity of recombinant protein drugs

Titelangaben

Canarelli, Stéphane ; Fisch, Igor ; Freitag, Ruth:
Hyphenation of multi-dimensional chromatography and mass spectrometry for the at-line-analysis of the integrity of recombinant protein drugs.
In: Journal of Chromatography B. Bd. 775 (Juli 2002) Heft 1 . - S. 27-35.
ISSN 1873-376X
DOI: https://doi.org/10.1016/S1570-0232(02)00286-6

Volltext

Link zum Volltext (externe URL): Volltext

Abstract

A robust tool is proposed for the rapid at-line verification of the identity and integrity of (recombinant) proteins, namely the hyphenation of multidimensional chromatography and mass spectrometry (MS). A recombinant human antibody produced in Chinese hamster ovary cells is taken as pertinent example. The recombinant human antibody is first captured from the production environment by affinity chromatography (rProtein A, isolation/concentration of the target molecule) and automatically transferred to an enzyme reactor (immobilized trypsin column) for digestion, thereby yielding different peptides corresponding to the protein sequence. The peptides are then separated on a reversed-phase column before being analyzed and identified by MS. This step does not require a fine resolution since the mass spectrometer can identify a variety of substances at the same time. The results are then analyzed in silico with suitable bio-informatic tools. When the gene sequence of the protein product is known, proteolytic cleavages can be predicted and the exact mass and hence the amino acid sequence of each peptide can thereby be deduced. Fitting experimental data and reference peptide sequences then provides important information about the integrity of the protein and more particularly about its sequence. In our case, the integrity of 45 of the light and 75 of the heavy chain sequences of the antibody could be verified within minutes.

Weitere Angaben

Publikationsform: Artikel in einer Zeitschrift
Begutachteter Beitrag: Ja
Keywords: Recombinant protein drugs
Institutionen der Universität: Fakultäten > Fakultät für Ingenieurwissenschaften > Lehrstuhl Bioprozesstechnik > Lehrstuhl Bioprozesstechnik - Univ.-Prof. Dr. Ruth Freitag
Fakultäten
Fakultäten > Fakultät für Ingenieurwissenschaften
Fakultäten > Fakultät für Ingenieurwissenschaften > Lehrstuhl Bioprozesstechnik
Titel an der UBT entstanden: Nein
Themengebiete aus DDC: 500 Naturwissenschaften und Mathematik
500 Naturwissenschaften und Mathematik > 500 Naturwissenschaften
600 Technik, Medizin, angewandte Wissenschaften
600 Technik, Medizin, angewandte Wissenschaften > 620 Ingenieurwissenschaften
Eingestellt am: 25 Feb 2016 15:17
Letzte Änderung: 25 Feb 2016 15:17
URI: https://eref.uni-bayreuth.de/id/eprint/31068