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Transcript
Quality Control of Intact Recombinant Proteins using
Sensitive High-Resolution Mass Spectrometry
Arnd Ingendoh, Dirk Wunderlich, Christian Albers
Bruker Daltonik GmbH, Bremen, Germany
The production of recombinant proteins is one of the fastest growing sectors in the pharmaceutical
industry as these proteins are increasingly used as drugs. With this interest in new
biopharmaceuticals proper quality control is needed to ensure the use of the right batches in the
proteins production. This includes knowledge about the correct amino acid sequence as well as
characterization of modification sites. Using a high-resolution instrument is recommended due to
the heterogeneity of those samples as well as the advantage of having an exact mass on the intact
protein and enzymatic fragments.
We use an ultrahigh-resolution QTOF for the LC/MS analysis of various proteins, like recombinant
IgG. The IgG was obtained from Chinese Hamster Ovary (CHO) cells. In addition, a mixture of
recombinant proteins was obtained from E.Coli cells. Proteins were separated with a Zorbax SBC8,
Rapid Resolution Cartridge (2.1 x 30 mm, 3.5 µm) within 15 minutes and analyzed by MS.
The resolution and wide mass range of the instrumental setup allowed for discrimination of discrete
changes in the glycosylation patterns even for proteins like IgG. Further analysis of the recombinant
IgG after reduction and alkylation allowed to measure the MW of the light chain at 25 kDa with 0.2
ppm mass accuracy. The MS spectra of the recombinant E. coli proteins allowed to distinguish
between protein monomers and dimers, covering a mass range up to 40 kDa.
The data was processed by specially designed software modules, which compare mass patterns and
accuracies with expected values ensuring a fast and reliable information retrieval, which is
mandatory for quality control. In combination with the possibility of running the intact proteins
directly, a high-throughput analysis is possible.
Organized and Produced by:
www.isranalytica.org.il
P.O.B 4034 Ness-Ziona 70400, Israel
Tel: +972-8-931-3070, Fax: +972-8-931-3071
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