Intact mass analysis was performed as described previously.27 (link),28 (link) Briefly, samples were resuspended in 2% acetonitrile and 0.1% trifluoroacetic acid and loaded onto a Jupiter 300 C5 column (Phenomenex) using an Agilent 1200 high-performance liquid chromatography (HPLC). Five micrograms of the GBSIa-EPAComPΔ28, GBSIb-EPAComPΔ28, and GBSIII-EPAComPΔ28 bioconjugate samples were desalted by washing with 2% acetonitrile ad 0.1% formic acid for 2 min at a flow rate of 0.25 mL/min and then separated using a linear gradient of 80% acetonitrile and 0.1% formic acid (2–80% acetonitrile over 12 min using 0.25 mL/min). Samples were infused into a 6520 Accurate mass Q-TOF mass spectrometer (Agilent), and MS1 mass spectra were acquired at 1 Hz between a mass range of 300 and 3000 m/z. Intact mass analysis and deconvolution were performed using MassHunter B.06.00 (Agilent).
Jupiter 300 c5 column
Manufactured by Phenomenex
Sourced in United States
The Jupiter 300 C5 column is a reversed-phase high-performance liquid chromatography (HPLC) column designed for the separation and analysis of a wide range of analytes. It features a 5-micron particle size and a 300-angstrom pore size, providing efficient and high-resolution separations. The column is constructed with a C5 stationary phase, which offers selectivity and retention characteristics suitable for a diverse set of compounds.
Automatically generated - may contain errors
Lab products found in correlation
Masshunter b 06, by Agilent Technologies (3 mentions)
6520 accurate mass q tof mass spectrometer, by Agilent Technologies (2 mentions)
Agilent 1200 infinity hplc, by Agilent Technologies (1 mentions)
6520 accurate mass quadrupole time of flight mass spectrometer, by Agilent Technologies (1 mentions)
Acquity uplc h class system, by Waters Corporation (1 mentions)
Xevo g2 xs qtof quadrupole time of flight mass spectrometer, by Waters Corporation (1 mentions)
4 protocols using jupiter 300 c5 column
1
Intact Mass Analysis of Bioconjugates
2
Intact Mass Analysis of Bioconjugates
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Intact mass analysis was performed on a 6520 Accurate Mass Quadrupole Time-of-Flight Mass Spectrometer coupled to an Agilent 1200 Infinity HPLC (Agilent, Santa Clara, CA, USA) using a Jupiter 300 C5 column (2mm*50mm, Phenomenex). Protein samples were resuspended in 2% acetonitrile, 0.1% trifluoroacetic acid and loaded/separated on the C5 column at a flow rate of 0.25 mL/min. 2 to 5 μg of each bioconjugate was desalted on column for 2 min with Buffer A (2% acetonitrile 0.1% formic acid) before being separated by altering the percentage of Buffer B (80% acetonitrile, 0.1% formic acid) from 0% to 100% over 16.5 min. The column was then held at 100% Buffer B for 0.5 min before being equilibrated for 1 min with Buffer A, for a total run time of 20.0 min. Samples were infused into the Time-of-Flight Mass Spectrometer using electrospray ionization (ESI) and MS1 mass spectra acquired with a mass range of 300–3000m/z at 1 Hz. Intact mass analysis and deconvolution was performed using MassHunter B.06.00 (Agilent).
3
Intact Mass Analysis of GBS Bioconjugates
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Intact mass
analysis was performed
as described previously.27 (link),28 (link) Briefly, samples were
resuspended in 2% acetonitrile and 0.1% trifluoroacetic acid and loaded
onto a Jupiter 300 C5 column (Phenomenex) using an Agilent 1200 high-performance
liquid chromatography (HPLC). Five micrograms of the GBSIa-EPAComPΔ28, GBSIb-EPAComPΔ28, and GBSIII-EPAComPΔ28 bioconjugate samples were desalted by washing
with 2% acetonitrile ad 0.1% formic acid for 2 min at a flow rate
of 0.25 mL/min and then separated using a linear gradient of 80% acetonitrile
and 0.1% formic acid (2–80% acetonitrile over 12 min using
0.25 mL/min). Samples were infused into a 6520 Accurate mass Q-TOF
mass spectrometer (Agilent), and MS1 mass spectra were acquired at
1 Hz between a mass range of 300 and 3000 m/z. Intact mass analysis and deconvolution were performed
using MassHunter B.06.00 (Agilent).
analysis was performed
as described previously.27 (link),28 (link) Briefly, samples were
resuspended in 2% acetonitrile and 0.1% trifluoroacetic acid and loaded
onto a Jupiter 300 C5 column (Phenomenex) using an Agilent 1200 high-performance
liquid chromatography (HPLC). Five micrograms of the GBSIa-EPAComPΔ28, GBSIb-EPAComPΔ28, and GBSIII-EPAComPΔ28 bioconjugate samples were desalted by washing
with 2% acetonitrile ad 0.1% formic acid for 2 min at a flow rate
of 0.25 mL/min and then separated using a linear gradient of 80% acetonitrile
and 0.1% formic acid (2–80% acetonitrile over 12 min using
0.25 mL/min). Samples were infused into a 6520 Accurate mass Q-TOF
mass spectrometer (Agilent), and MS1 mass spectra were acquired at
1 Hz between a mass range of 300 and 3000 m/z. Intact mass analysis and deconvolution were performed
using MassHunter B.06.00 (Agilent).
4
Intact Mass Analysis of Bioconjugates
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Intact mass analysis was performed on a Xevo G2-XS QTof Quadrupole Time-of-Flight Mass Spectrometer coupled to an ACQUITY UPLC H-class system (Waters) using a Jupiter 300 C5 column (2mm*50mm, Phenomenex). Protein samples were resuspended in 2% acetonitrile, 0.1% trifluoroacetic acid and loaded/separated on the C5 column at a flow rate of 0.25 mL/min. 2 μg of each bioconjugate (K2-EPA and O1-EPA) was desalted on column for 2 min with Buffer A (2% acetonitrile 0.1% formic acid) before being separated by altering the percentage of Buffer B (80% acetonitrile, 0.1% formic acid) from 0% to 100% over 10 min. The column was then held at 100% Buffer B for 0.5 min before being equilibrated for 1 min with Buffer A, for a total run time of 13.5 min. Samples were infused into the Xevo G2-XS QTof Quadrupole Time-of-Flight Mass Spectrometer using electrospray ionization (ESI) and MS1 mass spectra acquired with a mass range of 400–2000m/z at 1 Hz. Scans across the apex of the elution peaks were summed then peak lists exported. Deconvolution of proteoforms within K2-EPA and O1-EPA were undertaken using UniDec [54 (link)].
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