This application note describes a method for the rapid separation of recombinant human erythropoietin using the Agilent ZORBAX 300SB-C18 1.8 µm Rapid Resolution High Definition (RRHD) column. This method has increased sensitivity, reduced separation time and improved run-to-run reproducibility, which is in great demand by biotechnology and pharmaceutical companies. The C18 StableBond coating technology provides increased sensitivity, exceptional pH, and thermal stability. The use of a 1.8-µm column designed for UHPLC systems significantly reduces analysis time, critical for increasing the efficiency of QC for protein primary structure analysis. The data presented here focus on the fast separation of various recombinant human erythropoietin isoforms. Like many other biomolecules, recombinant EPO protein exhibits heterogeneity due to modifications that occur during manufacturing. Methods are optimized for flow rates, gradient and reproducibility under acidic conditions that contain trifluoroacetic acid.
This document presents an application note on the fast separation of recombinant human erythropoietin (rEPO) using the Agilent ZORBAX RRHD 300SB-C18 1.8 um column. The study highlights the column's ability to provide increased sensitivity, reduced separation time, and improved run-to-run reproducibility for protein analysis.
| Column dimensions | 2.1 x 50 mm |
|---|---|
| Particle size | 1.8 um |
| Maximum pressure | 1,200 bar |
| Temperature stability | 90 °C |
| Sample concentration | 1.0 mg/mL |
| Injection volume | 3 uL |
| Flow rate | 1.0 mL/min |
| Detector wavelength | 280 nm |
The system operates at a maximum pressure of 1,200 bar.
The column used has a particle size of 1.8 um.
Yes, the columns are stable up to 90 °C.