Proteins were diluted to 1 µM with 0.1% (v/v) formic acid and injected onto a C4 BEH 1.7 µm, 1.0 × 100 mm, UPLC column using the Acquity I class LC (Waters) system. Proteins were eluted with a 15 min gradient of acetonitrile (2% (v/v) to 80% (v/v)) in 0.1% (v/v) formic acid using a flow rate of 50 µl min−1. The analytical column outlet was directly interfaced through an electrospray ionization source, with a time-of-flight (TOF) mass spectrometer (BioAccord, Waters). Data were acquired over a m/z range of 300–8,000, in positive-ion mode with a cone voltage of 40 V. Scans were summed together manually and deconvoluted using MaxEnt1 (Masslynx, Waters). The parameters used were as follows; input m/z range (Da): 600–2,000; output mass range (Da): 30000–60000; TOF resolution: 10000.00; and iterate to convergence.
Bioaccord
Manufactured by Waters Corporation
Sourced in United States
The BioAccord is a liquid chromatography-mass spectrometry (LC-MS) system designed for routine analysis of biotherapeutics and biomolecules. It provides high-resolution, accurate-mass data to support characterization and quantitation of complex biomolecules.
Automatically generated - may contain errors
2 protocols using bioaccord
1
Protein Analysis by LC-MS with TOF
2
Characterization of Chemical Compounds
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General Methods Physical data were measured as follows: 1 H, 13 (link) C, and 31 P-NMR spectra were recorded at 400 or 500 MHz, 100 or 125 MHz, and 162 or 202 MHz instruments (Bruker FT-NMR AV400 or AV500), respectively in CDCl 3 or dimethyl sulfoxide (DMSO)-d 6 as the solvent with tetramethylsilane or H 3 PO 4 as an internal or external standard. Chemical shifts are reported in parts per million (ppm), and signals are expressed as s (singlet), d (doublet), t (triplet), m (multiplet), or br (broad). All exchangeable protons were detected by addition of D 2 O. Mass spectra were measured on a SQD2 (Waters, U.S.A.) and BioAccord (Waters). TLC was done on Merck Kieselgel F254 precoated plates. Silica gels used for column chromatography were KANTO Chemical silica gel 60 and KANTO Chemical silica gel 60N (neutral).
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