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Poroshell 300sb c8 column

Manufactured by Agilent Technologies
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The Poroshell 300SB-C8 column is a high-performance liquid chromatography (HPLC) column designed for the separation and analysis of a wide range of compounds. The column features a silica-based stationary phase with octylsilyl (C8) functional groups, providing effective separation capabilities for both polar and non-polar analytes. The Poroshell technology employed in this column offers increased column efficiency and reduced analysis time compared to traditional HPLC columns.

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Lab products found in correlation

Exactive plus orbitrap mass spectrometer, by Thermo Fisher Scientific (2 mentions) Lxq system, by Thermo Fisher Scientific (1 mentions) Sb c8 column, by Agilent Technologies (1 mentions) Qualitative analysis software, by Agilent Technologies (1 mentions) 1100 lc system, by Agilent Technologies (1 mentions) Hi plex ca column, by Agilent Technologies (1 mentions) 1260 infinity, by Agilent Technologies (1 mentions) 1290 infinity 2 hplc, by Agilent Technologies (1 mentions)

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Poroshell 300SB-C8 300SB-C8 column 300SB-C8 C8 column

6 protocols using poroshell 300sb c8 column

1

LC-ESI-MS Characterization of Fc Glycosylation

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LC-ESI-MS was
performed on a LXQ system (Thermo Scientific) with
an Agilent Poroshell 300SB-C8 column (5 μm, 75 × 1 mm).
Fc samples were reduced in 40 mM tris (2-carboxyethyl) phosphine at
37 °C for 20 min before LC-ESI-MS measurement. LC separation
of the resulting monomers was performed at 40 °C eluting with
a linear gradient of 20–40% acetonitrile containing 0.1% formic
acid within 10 min at a flow rate of 0.25 mL/min. Percent glycan transfer
was determined by comparing mass spectral abundances of oxime-GlcNAc
Fc2 and oxime-(S2/G2) Fc2.
Smith E.L., Giddens J.P., Iavarone A.T., Godula K., Wang L.X, & Bertozzi C.R. (2014). Chemoenzymatic Fc Glycosylation via Engineered Aldehyde Tags. Bioconjugate Chemistry, 25(4), 788-795.
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2

LC-TOF-MS Analysis of FGF2 and IGF1

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Enriched FGF2 and IGF1 samples were analyzed at the Mayo Proteomics Core Facility using liquid chromatography separation combined with a time-of-flight mass spectrometry (LC-TOF-MS). Injections of 8 µL of recombinant protein solution (20 ng/µL) were loaded onto an Agilent Poroshell 300SB-C8 column (1 × 75 mm, 5 µm) or Zorbax SB-C8 column (2.1 × 50 mm, 3.5 µm) on an Agilent 1100 LC system. Separation was achieved using a linear gradient of H2O, acetonitrile, and 0.1% formic acid. Mass spectrometry was performed on an Agilent MSD/TOF system using electrospray ionization in positive ion mode. The mass spectrometry acquisition method was set to acquire data over the entire duration of each LC gradient, in the mass range from 400 to 2500 m/z. Chromatograms and raw spectra were produced by Agilent Qualitative Analysis Software. Deconvoluted spectra of molecular mass were generated from Agilent Protein BioConfirm Software.
Park S., Arrell D.K., Reyes S., Park E.Y, & Terzic A. (2017). Conventional and unconventional secretory proteins expressed with silkworm bombyxin signal peptide display functional fidelity. Scientific Reports, 7, 14499.
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3

HPLC Separation of Protein and Sugars

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The composition of particles was determined using a high-performance liquid chromatography (HPLC) instrument (1260 Infinity by Agilent) equipped with an evaporative light scattering detector. The HPLC method used was based on separating BSA from lactose and glycerol using an Ultracel-30K Amicon Ultra centrifugal filter (Millipore) and then running them on two different columns based on reversed phase chromatography using a Poroshell 300SB-C8 column (Agilent) for the BSA in the mixture and ion exchange chromatography using a Hi-Plex Ca column (Agilent) for lactose and glycerol (Table S1, Supporting Information).
Khodabandehlou K., Kumbhar A.S., Habibi S., Pandya A.A., Luft J.C., Khan S.A, & DeSimone J.M. (2015). Silylated Precision Particles for Controlled Release of Proteins. ACS applied materials & interfaces, 7(10), 5756-5767.
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4

Orbitrap-based LC-ESI-MS Metabolite Analysis

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LC–ESI–MS analysis was performed with an Exactive Plus Orbitrap Mass Spectrometer (Thermo Scientific) equipped with an Agilent Poroshell 300SB C8 column (5 μm, 1.0 × 75 mm) with a gradient elution of 25−35% aq MeCN containing 0.1% FA for 6 min, 0.4 mL/min. Mass spectra were deconvoluted using MagTran (ver 1.03 b2).
Kao K.S., Gupta A., Zong G., Li C., Kerschbaumer I., Borghi S., Achkar J.M., Bournazos S., Wang L.X, & Ravetch J.V. (2022). Synthetic nanobodies as tools to distinguish IgG Fc glycoforms. Proceedings of the National Academy of Sciences of the United States of America, 119(48), e2212658119.
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5

Antibody Analysis by Mass Spectrometry

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The antibody samples in PBS were incubated with IdeS at 37 °C for 2 h. The samples were analyzed by with Exactive Plus Orbitrap Mass Spectrometer (Thermo Scientific) equipped with an Agilent Poroshell 300SB C8 column (5 μm, 1.0 × 75 mm) with gradient elution of water containing 0.1% formic acid as phase A, MeCN containing 0.1% formic acid as phase B. Mass spectra were deconvoluted using MagTran (ver 1.03 b2).
Ou C., Prabhu S.K., Zhang X., Zong G., Yang Q, & Wang L.X. (2022). Synthetic antibody-rhamnose cluster conjugates show potent complement-dependent cell killing by recruiting natural antibodies. Chemistry (Weinheim an der Bergstrasse, Germany), 28(16), e202200146.
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6

Quantitative LC/MS Protein Analysis

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LC/MS analysis was performed on an Agilent 1290 Infinity II HPLC connected to an Agilent 6530B QTOF AJS-ESI. 1 μg of protein was injected onto a Poroshell 300SB-C8 column (2.1 x 75 mm, 5-Micron, room temperature, Agilent) using a linear gradient from 5 to 75% acetonitrile over 9.5 minutes with 0.1% formic acid as the aqueous phase after an initial hold at 5% acetonitrile for 0.5 min (0.4 mL/min). The following parameters were used during acquisition:
Fragmentor voltage 175 V, gas temperature 300ºC, gas flow 12 L/min, sheath gas temperature 350ºC, sheath gas flow 12 L/min, nebulizer pressure 35 psi, skimmer voltage 65 V, Vcap 5000 V, 3 spectra/s. Intact protein masses were obtained via deconvolution using the Maximum Entropy algorithm in Mass Hunter Bioconfirm (V10, Agilent).
Santiago S., Ad O., Shah B., Zhang Z., Zhang X., Chatterjee A., & Schepartz A. (2021). Genetic code expansion in the engineered organism Vmax X2: High yield and exceptional fidelity.
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