This application note describes how to use Agilent Bio IEX (WCX and SCX) and MAb nonporous 3 µm poly(styrene divinylbenzene) columns to separate proteins.
This document presents a method for protein separation using Agilent Bio IEX (WCX and SCX) and MAb nonporous, 3 µm poly(styrene divinylbenzene) columns. It compares the elution differences between these column types when separating Cytochrome c, ribonuclease A, and lysozyme.
| Column Type | Agilent Bio IEX (WCX, SCX) and MAb |
|---|---|
| Particle Size | 3 µm |
| Column Dimensions | 4.6 x 50 mm |
| Flow Rate | 0.5 mL/min |
| Temperature | Ambient |
| Injection Volume | 2 µL |
| Detection Wavelength | 220 nm |
| Instrument | Agilent Infinity 1260 Bio-inert HPLC system |
The test used Cytochrome c, ribonuclease A, lysozyme, and a protein mix.
The columns used have a particle size of 3 µm.
WCX and MAb elute in the order cytochrome c, RNase, then lysozyme, while SCX elutes RNase A, cytochrome c, then lysozyme.