The 1H and 13C- NMR spectra were obtained on a Bruker Avance 400 and 500 MHz in deuterated chloroform (CDCl3). Chemical shifts are in δ units (ppm) with TMS (0.0 ppm). PEG conjugates were purified using HPLC on the Hewlett- Packard liquid chromatography system Series 1100. Ultraviolet-Visible detection was performed using HP photodiode array detector. For purification and analysis, Waters X-Bridge BEHC4 column (4.6 mm × 100 mm, 10K-500K) (analytical column) was used with a flow rate at 1.0 ml/min, at 80°C and was monitored at 280 nm as well as Agilent Zorbax 300SB-C18 4.6 × 250 mm (Semi-Prep column) at 3.0 ml / min at 45 °C and was monitored at 280 nm. MALDI-ToF measurements were performed on Bruker Autoflex MALDI-ToF. BD FACSVerse flow cytometer was used for flow cytometry work.
X bridge behc4 column
Manufactured by Waters Corporation
Sourced in United States
The X-Bridge BEHC4 column is a reversed-phase high-performance liquid chromatography (HPLC) column designed for the separation and analysis of a wide range of organic compounds. The column features a hybrid organic-inorganic silica-based stationary phase that provides high efficiency, excellent peak shape, and enhanced chemical and thermal stability.
Automatically generated - may contain errors
Lab products found in correlation
Series 1100, by Hewlett-Packard (2 mentions)
Autoflex maldi tof, by BD (2 mentions)
Zorbax 300sb c18, by Agilent Technologies (2 mentions)
Avance 400, by Bruker (2 mentions)
Facsverse flow cytometer, by BD (2 mentions)
E2695 separations module, by Waters Corporation (1 mentions)
Uhplc system, by Agilent Technologies (1 mentions)
4 protocols using x bridge behc4 column
1
Characterization and Purification of PEG Conjugates
2
Characterization and Purification of PEG Conjugates
3
Protein Profiling and SDS-PAGE Analysis
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The protein profiles of permeate were determined using a Waters e2695 Separations Module (Waters Corp., Milford, MA, USA), equipped with a Waters XBridge BEH C4 column (250 mm × 4.6 mm 179 I.D.) and a Waters 2489 UV/Visible detector at 220 nm according to the method of Visser et al. [27 (link)]. The protein contents of permeate were calculated as a percentage of their peak area chromatogram relative to a BRM milk sample chromatogram.
A reducing SDS-page was conducted. Samples were diluted by a factor of 8 and then mixed with a loading buffer (25 mM Tris-HCl, 100 g/L glycerol (v/v), 20 g/L SDS (w/v), 50 g/L β-Mercaptoethanol (v/v) and 1 g/L bromophenol blue (w/v)) at 1:1. The mixed samples (10 μL) were loaded onto an SDS-PAGE gel (40 g/L acrylamide stacking and a 130 g/L acrylamide separating gel). The procedure of running, staining and de-staining for gels was completed according to Verdi et al. [28 (link)].
A reducing SDS-page was conducted. Samples were diluted by a factor of 8 and then mixed with a loading buffer (25 mM Tris-HCl, 100 g/L glycerol (v/v), 20 g/L SDS (w/v), 50 g/L β-Mercaptoethanol (v/v) and 1 g/L bromophenol blue (w/v)) at 1:1. The mixed samples (10 μL) were loaded onto an SDS-PAGE gel (40 g/L acrylamide stacking and a 130 g/L acrylamide separating gel). The procedure of running, staining and de-staining for gels was completed according to Verdi et al. [28 (link)].
4
UHPLC Analysis of API Content
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API content in drug product formulations was determined using Agilent UHPLC system equipped with a binary pump. 5 mL of sample solution in 30/70 THF/water (v/v) was injected on to a Waters XBridge BEH C4 column (50 mm x 4.6 mm i.d., 3.5 mm, 300 A ) maintained at 70 °C. Aqueous mobile phase (MPA) was composed of 0.1% TFA in milli-Q water and organic mobile phase (MPB) was 0.1% TFA in acetonitrile. Flow rate was set to 0.5 mL/min. Gradient program was
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