Differentiation of isomeric N-glycan structures by normal-phase liquid chromatography-MALDI-TOF/TOF tandem mass spectrometry

The detailed characterization of protein N-glycosylation is very demanding given the many different glycoforms and structural isomers that can exist on glycoproteins. Here we report a fast and sensitive method for the extensive structure elucidation of reducing-end labeled N-glycan mixtures using a combination of capillary normal-phase HPLC coupled off-line to matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) and TOF/TOF-MS/MS. Using this method, isobaric N-glycans released from honey bee phospholipase A2 and Arabidopsis thaliana glycoproteins were separated by normal-phase chromatography and subsequently identified by key fragment ions in the MALDI-TOF/TOF tandem mass spectra. In addition, linkage and branching information were provided by abundant cross-ring and "elimination" fragment ions in the MALDI-CID spectra that gave extensive structural information. Furthermore, the fragmentation characteristics of N-glycans reductively aminated with 2-aminobenzoic acid and 2-aminobenzamide were compared. The identification of N-glycans containing 3-linked core fucose was facilitated by distinctive ions present only in the MALDI-CID spectra of 2-aminobenzoic acid-labeled oligosaccharides. To our knowledge, this is the first MS/MS-based technique that allows confident identification of N-glycans containing 3-linked core fucose, which is a major allergenic determinant on insect and plant glycoproteins.

Identifying the knowledge management structures in enterprise systems projects

Enterprise System (ES) implementation and management are knowledge intensive tasks that inevitably draw upon the experience of a wide range of people with diverse knowledge capabilities. Knowledge Management (KM) has been identified as a critical success factor in ES projects. Despite the recognized importance of managing knowledge for ES benefits realization, systematic attempts to conceptualize KM-structures have been few. Where the adequacy of KM-structures is assessed, the process and measures are typically idiosyncratic and lack credibility. Using the ‘KM-process’, itself based in sociology of knowledge, this paper conceptualizes four main constructs to measure the adequacy of KM-structures. The SEM model is tested using 310 responses gathered from 27 ES installations that had implemented SAP R/3. The findings reveal six constructs for KM-structure. Furthermore, the paper demonstrates the application of KM-structures in the context of ES using the Adaptive Structuration Theory. The results demonstrate that having adequate KM-structures in place, while necessary, is not sufficient. These rules and resources must be appropriated to have greater positive influence on the Enterprise System. Furthermore, the study provides empirical support for knowledge-based theory by illustrating the importance of knowledge use/re-use (vs. knowledge creation) as the most important driver in the process of KM.