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catalog description
| Note: Because the capillary wall is populated with Si-H, where not covered with cholesterol basic proteins are not
strongly adsorbed and analyses are very reproducible. |
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CelerityCE™ Application Sheet
PEGylated-Protein Profiling for Batch Consistency
Versatile and reproducible technique for the analysis of biomolecules
Method Conditions:
Voltage: 20kV
Capillary: CelerityCE™ Cholesterol, i.d. 50µm, total length 50cm, effective length 41.5cm
Catalog No: 04925-50
Injection: 3 seconds @ 50 mbar
Stock Buffer: 0.3mol/L phosphoric acid and 0.19 mol/L Tris[tris(hydroxymethyl)aminomethane], pH=2.14
Run Buffer: Stock buffer diluted to 1:10, pH=2.14
Sample: PEG-catalase
Detection: UV @ 210nm
Instrument: Agilent 3D HPCE |
Attachment of polyethylene glycol (PEG) moieties to proteins increases solubility, lowers toxicity and the dosage due to
longer residence time in the body and does not cause loss of biological activity of therapeutic agents. A complex,
reproducible profile of PEG-catalase was obtained using Capillary Electrophoresis with cholesterol coated capillaries,
employing the hybrid technique known as Open Tubular Capillary Electro-Chromatography, which combines the separation
mechanism found in both chromatographic (solute/bonded phase interaction) and electrophoretic (differences in
electrophoretic mobility) methods.
Discussion:
When the sample was injected onto a bare fused silica capillary only random non-reproducible noise spikes were observed.
Using CelerityCE capillaries is an effective method for verifying the purity of PEG-proteins or for establishing
batch-to-batch reproducibility. High resolution of the spectra makes the method comparable to MALDI-TOF-MS but is much
less costly.
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