Human nasal IgA and recombinant IgG and IgA antibodies were subjected to glycan analysis. Human nasal IgA was purified from nasal wash concentrates using CaptureSelect IgA (Thermo Fisher Scientific), according to the manufacturer’s instructions. Purified antibodies were concentrated using Amicon Ultracell (Merck, Darmstadt, Germany) centrifugation units with a cut-off of 30 kDa. The human nasal wash concentrate samples used here were residual samples collected from a past clinical study [60 (link)]. Antibodies were deglycosylated by PNGase F (Takara Bio, Shiga, Japan) treatment. The released N-glycans were purified and labeled with 2-aminobenzamide using BlotGlyco (Sumitomo Bakelite, Tokyo, Japan), according to the manufacturer's instructions. Liquid chromatography-mass spectrometry (LC-MS) analysis was performed by Sumitomo Bakelite using a LC-MS-IT-TOF apparatus (Shimadzu, Kyoto, Japan). N-glycan structures were predicted using the GlycoMod Tool (http://web.expasy.org/glycomod/ ) [61 (link)].
Lc ms it tof apparatus
Manufactured by Shimadzu
Sourced in Japan
The LC-MS-IT-TOF apparatus is a highly specialized analytical instrument that combines liquid chromatography (LC) with mass spectrometry (MS) using an ion trap (IT) and time-of-flight (TOF) mass analyzer. This device is designed to provide precise and accurate analysis of complex samples, allowing for the identification and quantification of a wide range of chemical compounds.
Automatically generated - may contain errors
Lab products found in correlation
Pngase f, by Takara Bio (1 mentions)
Amicon ultracell, by Merck Group (1 mentions)
Captureselect iga, by Thermo Fisher Scientific (1 mentions)
Blotglyco, by Sumitomo Bakelite (1 mentions)
A1 laser scanning confocal microscope, by Nikon (1 mentions)
F 7000 uorescence spectrometer, by Hitachi (1 mentions)
Gemini a ultra diffractometer, by Agilent Technologies (1 mentions)
Av 400 nmr spectrometer, by Bruker (1 mentions)
2 protocols using lc ms it tof apparatus
1
Glycan Profiling of Nasal IgA and Recombinant Antibodies
2
Synthesis and Characterization of a Fluorescent Coumarin Probe
Check if the same lab product or an alternative is used in the 5 most similar protocols
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All chemicals were obtained from commercial suppliers and directly used without further puri cation. 7-(diethylamino)coumarin-3-carbohydrazide was prepared according to the literature method [16] .
Analytical grade acetonitrile and deionized water were used as solvents for spectral measurements. 1 H NMR and 13 C NMR spectra were recorded on a Bruker Av400 NMR spectrometer. HRMS spectra were performed on a Shimadzu LCMS-IT-TOF apparatus. Fluorescence spectra were taken on a Hitachi F-7000 uorescence spectrometer. The X-ray diffraction data was collected on an Agilent Gemini A Ultra diffractometer (MoKα, λ = 0.71073 Å). Images of cells were obtained using a Nikon A1 confocal laser scanning microscope.
Analytical grade acetonitrile and deionized water were used as solvents for spectral measurements. 1 H NMR and 13 C NMR spectra were recorded on a Bruker Av400 NMR spectrometer. HRMS spectra were performed on a Shimadzu LCMS-IT-TOF apparatus. Fluorescence spectra were taken on a Hitachi F-7000 uorescence spectrometer. The X-ray diffraction data was collected on an Agilent Gemini A Ultra diffractometer (MoKα, λ = 0.71073 Å). Images of cells were obtained using a Nikon A1 confocal laser scanning microscope.
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