title: The characterization and quantitation of glycomic changes in CHO cells during a bioreactor campaign.
abstract: Within the biotechnology industry there is a continuous drive to better and more fully understand the biopharmaceutical process in order to achieve better process control. A method to monitor and quantitate glycomic changes that occur in CHO cells during a bioreactor campaign could help to address this. The goal of the method presented here is to provide data that may help to understand the changes in glycosylation that are occurring, within the cell, to proteins other than the expressed biotherapeutic. The method involves the lysing of cells to gain access to intracellular proteins. The expressed biotherapeutic is specifically removed by affinity chromatography, while the remaining proteins are subjected to deglycosylation by treatment with PNGase F. The released glycans are derivatized with isotopic tags, and quantitative analysis by MALDI-TOF MS is performed. The MALDI-TOF MS method allows for the simultaneous analysis of both neutral and sialylated glycans, displays a linear dynamic range over two orders of magnitude for both neutral and sialylated glycans and achieves sub-picomolar sensitivity. This method may yield valuable information that gives further insight into the inner-workings of CHO cells, potentially taking another step towards fully understanding and controlling the biopharmaceutical process.
authors: Biogen Idec, 14 Cambridge Center, Cambridge, Massachusetts 02142, USA. samnang.tep@biogenidec.com
authors: Samnang
authors: S
authors: Tep
authors: Marina
authors: M
authors: Hincapie
authors: William S
authors: WS
authors: Hancock
mesh_terms: D000818:Animals; D019149:Bioreactors; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D004591:Electrophoresis, Polyacrylamide Gel; D054794:Glycomics / Q000379:methods*; D006023:Glycoproteins / Q000032:analysis / Q000378:metabolism*; D006031:Glycosylation; D007553:Isotope Labeling; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase / Q000032:analysis / Q000378:metabolism; D011134:Polysaccharides / Q000032:analysis / Q000378:metabolism*; D015203:Reproducibility of Results; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013205:Staphylococcal Protein A / Q000032:analysis / Q000378:metabolism; D062367:ortho-Aminobenzoates / Q000737:chemistry / Q000378:metabolism
chemical_list: D006023:Glycoproteins; D011134:Polysaccharides; D013205:Staphylococcal Protein A; D062367:ortho-Aminobenzoates; C031385:anthranilic acid; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase