Final ProgramCPSA 2011
Science and Technology Coming Together to Make a Difference
October 3 - 6, 2011
Bucks County Sheraton Hotel
Langhorne, PA
Poster Abstract #27
Development of a Methodology for Rapid Characterization of Biological Therapeutics
1) Bruker Daltonics, Billerica, MA; 2) Bruker Daltonics, Fremont, CA; 3) Bruker Daltonik, Bremen, Germany; 4) Bruker Daltonics, Coventry, UK
By 2014, it is expected that six of the top world’s top ten best selling drugs will be biopharmaceuticals. The characterization of these products presents many challenges due to their high molecular weight, the heterogeneous nature of proteins, and the complexity of the production process. Subtle changes in the manufacturing steps can introduce unexpected and unwanted modifications to the product. Therefore, a solution is required for fast, reliable, and quantitative characterization of biopharmaceuticals in a high throughput, automated manner. Fortunately, many of the tools used in classical small molecule Qual/Quant workflows are applicable to biopharmaceutical analysis. We describe a method based on LC-MS technology, combining ultra-high-resolution chromatography (U-HPLC), ultra high-resolution mass spectrometry (UHR-TOF-MS), and newly developed software –BioPharma Compass–to provide complete automation of the characterization workflow.
Results
20 samples of human IgG1 were analyzed using an automated workflow where LC-MS data were acquired on a UHR-TOF mass spectrometer and the spectra analyzed with the Maximum Entropy Deconvolution algorithm. The resulting peak lists were qualitatively and quantitatively compared to a "gold" reference standard. Using this approach any differences in the samples compared to the reference were automatically highlighted. Results show the high spectral mass accuracy and resolution of the platform is clearly capable of determining small differences in mass and intensity indicating differences in glycosylation.
The peptide mapping workflow was tested by analyzing 96 tryptic digest of BSA and transferrin by LC-MS. Data were matched automatically matched to the theoretical protein digests considering possible modifications. Sequence coverage of up to 95% was obtained for most samples. Future work will investigate the use of alternative proteolytic enzymes to obtain full sequence coverage.
Return to program