All measurements with the poly(dT) ladder were acquired on an ACQUITY UPLC I-Class system (Waters Corporation, Milford, MA, USA) coupled with a VION IMS qTOF mass spectrometer (Waters Corporation, Milford, MA, USA), measurements with the 24 nt thiolated oligonucleotide were acquired on a BioAccord LC-MS System with ACQUITY Premier (Waters Corporation, Milford, MA, USA). Source parameters were set as: 100 V source offset, 120 °C source temperature, 400 °C desolvation temperature, 800 L/h (L/h) desolvation gas flow and 50 L/h cone gas flow. The scan range of full MS was set to m/z 500.00–2000.00 at a scan rate of 1 Hz (1 s−1). Data acquisition and processing were performed with UNIFITM (1.9.13.9 and 3.0.0.15). Calibration of MS data was achieved throughconstant infusion of leucine enkephalin calibration solution by Waters (SKU: 186006013) at a flow rate of 10 µL/min.
Bioaccord lc ms system
Manufactured by Waters Corporation
Sourced in United States
The BioAccord LC-MS System is a liquid chromatography-mass spectrometry (LC-MS) platform designed for biopharmaceutical analysis. It integrates Waters' ACQUITY PREMIER Columns and Empower Chromatography Data Software to provide high-resolution separation and sensitive mass detection. The system is capable of analyzing a wide range of biomolecules, including proteins, peptides, and oligonucleotides.
Automatically generated - may contain errors
Lab products found in correlation
Acquity uplc system, by Waters Corporation (2 mentions)
Acquity uplc 1 class system, by Waters Corporation (1 mentions)
Vion ims qtof mass spectrometer, by Waters Corporation (1 mentions)
Source 30q resin, by GE Healthcare (1 mentions)
Protein beh c4 column, by Waters Corporation (1 mentions)
Acquity uplc 1 class plus, by Waters Corporation (1 mentions)
Leucine enkephalin, by Waters Corporation (1 mentions)
Acquity uplc peptide beh c18 column, by Waters Corporation (1 mentions)
7 protocols using bioaccord lc ms system
1
Optimized UPLC-IMS-QTOF Mass Spectrometry
2
Conjugation of GLP1/FGF21 Fusion Protein
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Example 3
To a solution of GLP1/FGF21 fusion protein (75 mg) in NaOH or Tris buffer was added with CRM reagent (i.e. HOOC—(CH2)16-CO-gGlu-2XADO, HOOC—(CH2)16-CO-gGlu-2XADO-EDA-CO—CH2, or HOOC—(CH2)20-CO-gGlu-2XADO-EDA-CO—CH2) (6 mg or 12 mg) in organic solvent dropwise. The reaction was stirred at room temperature for 1 h. Then the product was applied to anion exchange chromatography using Source 30Q resin (GE healthcare).
The conjugated fusion proteins (as shown in Table 2) were detected and characterized by LC-MS method with Waters BioAccord LC-MS system, or by UPLC with Waters Acquity UPLC system, using conditions optimized for different conjugates, following the supplier's manuals.
3
Conjugation of GLP-1 Protein
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Example 3
To a solution of a GLP-1 protein in NaOH was added with CRM reagent (i.e. HOOC—(CH2)16-CO-gGlu-2XADO) in organic solvent dropwise. The reaction was stirred at room temperature for 1 h. Then the product was applied to reverse phase chromatography. This provided the compounds as listed in Table 1 as shown above.
The conjugated GLP-1 proteins were detected and characterized by LC-MS method with Waters BioAccord LC-MS system, or by UPLC with Waters Acquity UPLC system, using conditions optimized for different conjugates, following the supplier's manuals.
4
Structural Characterization of IgG N-Glycans
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For structural characterization of IgG N-glycans prepared with AdvanceBio Gly-X N-glycan Prep with InstantPC Kit, the released and labelled N-glycans were analysed by liquid chromatography-mass spectrometry (LC-MS) on a BioAccord LC-MS System (Waters, USA). Chromatographic conditions were the same as described above for the analysis of InstantPC labelled glycans. The RDa mass detector was used in-line via electrospray ionization in positive mode. The settings were as follows: scan range, 50–2000 m/z; capillary voltage, 1.5 kV; cone voltage, 45 V; desolvation temperature, 300°C; and sampling rate, 2 Hz. Acquired data were automatically processed using the UNIFI 1.9.9.3 Scientific Information System. For the glycan identification, MS1 sum spectra were generated around the retention times (RT) of the chromatographic peak and annotation of MS1 sum spectra were inferred from the m/z values and deduced using GlycoMod software (https://web.expasy.org/glycomod/ , accessed on the 7th of July 2021)26 (link) MS interpretation was based on previously reported structures19 and biosynthetic pathways.
5
Peptide Conjugation of Trastuzumab
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0.5 mL of 10 mg/mL trastuzumab solution (68 μM) in PBS was mixed with 3.4 μL of 30 mM peptide reagent at a molar ratio of 1:3, and incubated for 15 min. The samples were diluted with distilled water and subjected to ion-exchange chromatography on IEC SP-825 (8 μm, 8.0 × 75 mm, Shodex) equilibrated with 0.1 M sodium acetate-HCl buffer (pH 5.5). The samples were eluted with a NaCl gradient from 0 to 0.4 M for 30 min at a flow rate of 0.8 mL/min to separate the unmodified, monopeptide and dipeptide. The ion-exchange chromatograms were acquired by monitoring the fluorescence detector at excitation/emission wavelengths of 280/340 nm. After dialysis against the distilled water, the collected fractions were lyophilized. The final yield of the peptide conjugation is approximately 80%. The modification on trastuzumab was confirmed using LC–MS on BioAccord LC–MS System (Waters) connected with Protein BEH C4 Column (2.1 × 5 mm, Waters) which was in a column oven at 80 °C. The elution was done by acetonitrile gradient from 1 to 50% both containing 0.1% formic acid at a flow rate of 0.4 mL/min.
6
Antibody Tryptic Digest Analysis
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Five micrograms (5 μg) of tryptic digested antibody were analysed on a BioAccord LC-MS system (Waters Corporation, United States). The system configuration includes an ACQUITY UPLC I-Class PLUS with an ACQUITY RDa detector (a compact time-of-flight mass detector) controlled by the compliance-ready UNIFI Scientific Information System software platform (Waters Corporation, United States). Samples were separated using an ACQUITY UPLC Peptide BEH C18 column (130 Å, 1.7 μm, 2.1 mm × 100 mm, Waters Corporation, United States) at 65°C and 250 μL/min, with a 60 min gradient from 1 to 40% of 0.1% formic acid in acetonitrile (mobile phase B). 0.1% formic acid in LCMS water was used as mobile phase A. The RDa mass detector was operated in positive electrospray ionisation full scan with fragmentation acquisition mode (MSe) at 2 Hz acquisition rate with a mass range of 400–2,000 m/z. The capillary voltage was set at 1.2 kV, cone voltage at 30 V, and the desolvation temperature was kept at 350°C. Leucine Enkephalin (Waters Corporation, United States) was used as the LockSpray compound for real-time mass correction.
7
Structural Characterization of Seminal Plasma Glycans
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For structural characterization of glycans eluted in SPGP14, the released and labeled N-glycans of prepared in-house seminal plasma standard were analyzed by liquid chromatography-mass spectrometry (LC-MS) on a BioAccord LC-MS System (Waters). Chromatographic conditions were the same as described above. The RDa mass detector was used in-line via electrospray ionization in positive mode. The settings were as follows: scan range, 50–2000 m/z; capillary voltage, 1.5 kV; cone voltage, 45 V; desolvation temperature, 300 °C; and sampling rate, 2 Hz. Acquired data were automatically processed using the UNIFI 1.9.9.3 Scientific Information System. For the glycan identification, MS1 sum spectra were generated around the retention times (RT) of the chromatographic peak reported as SPGP14 (RT 12.99–13.33 min). Annotation of MS1 sum spectra was inferred from the m/z values and deduced using GlycoMod software (https://web.expasy.org/glycomod/ , accessed on 7 July 2021). MS interpretation was based on previously reported structures [15 (link)] and biosynthetic pathways.
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