[en] Although a detailed finite element (FE) model provides more precise results, a lumped-mass stick (LMS) model is preferred because of its simplicity and rapid computational time. However, the reliability of LMS models has been questioned especially for structures dominated by higher modes and seismic inputs. Normally, the natural frequencies and dynamic responses of a LMS model based on tributary area mass consideration are different from the results of the FE model. This study proposes a basic updating technique to overcome these discrepancies; the technique employs the identical modal response, D(t), to the detailed FE model. The parameter D(t) is a time variable function in the dynamic response composition and it depends on frequency and damping ratio for each mode, independent of the structure’s mode shapes. The identical response D(t) for each mode is obtained from the frequency adaptive LMS model; the adaptive LMS model which can provide identical modal frequencies as the detailed FE model. Theoretical backgrounds and formulations of the updating technique are proposed. To validate the updating technique, two types of structures (a symmetric straight column and an unsymmetric T-shaped structure) are considered. From the seismic response results including base shear and base moment, the updating technique considerably improves the seismic response accuracy of the tributary area-based LMS model.

[en] Highlights: • M6P glycosylation is a key factor for lysosomal enzyme targeting and efficacy of ERT. • M6P glycosylation on rhGAA, the only ERT for Pompe disease, was not fully reported. • This study investigated M6P glycosylation on rhGAA using LC-ESI-HCD-MS/MS. • All M6P glycans constitute 1.0% of the 78 glycans of rhGAA. • Four types of ten M6P glycans and their three attachment sites are newly identified. Myozyme is a recombinant human acid alpha-glucosidase (rhGAA) that is currently the only drug approved for treating Pompe disease, and its low efficacy means that a high dose is required. Mannose-6-phosphate (M6P) glycosylation on rhGAA is a key factor influencing lysosomal enzyme targeting and the efficacy of enzyme replacement therapy (ERT); however, its complex structure and relatively small quantity still remain to be characterized. This study investigated M6P glycosylation on rhGAA using liquid chromatography (LC)–electrospray ionization (ESI)–high-energy collisional dissociation (HCD) tandem mass spectrometry (MS/MS). The glycans released from rhGAA were labeled with procainamide to improve mass ionization efficiency and the sensitivity of MS/MS. The relative quantities (%) of 78 glycans were obtained, and 1.0% of them were glycans containing M6P (M6P glycans). These were categorized according to their structure into 4 types: 3 newly found ones, comprising high-mannose-type M6P glycans capped with N-acetylglucosamine (GlcNAc) (2 variants, 17.5%), hybrid-type M6P glycans (2 variants, 11.2%), and hybrid-type M6P glycans capped with GlcNAc (3 variants, 6.9%), as well as high-mannose-type M6P glycans (3 variants, 64.4%). HCD-MS/MS spectra identified six distinctive M6P-derived oxonium ions. The glycopeptides obtained from protease-digested rhGAA were analyzed using nano-LC-ESI-HCD-MS/MS, and the extracted-ion chromatograms of M6P-derived oxonium ions confirmed three M6P glycosylation sites comprising Asn 140, Asn 233 (newly found), and Asn 470 attached heterogeneously to nine M6P glycans (two types), eight M6P glycans (four types), and seven M6P glycans (two types), respectively. This is the first study of rhGAA to differentiate M6P glycans and identify their attachment sites, despite rhGAA already being an approved drug for Pompe disease.