In many cases protein purification requires two or even more steps to achieve sufficient purity. The transition from one step to another can be very time consuming and requests manual interaction. These limitations illustrate the need for FPLC systems enabling automated two step purification. This application highlights the possibilityof combining two subsequent chromatography protocols. This was achieved using the KNAUER Lab standard Multi Method FPLC system adapted for automatic two step purification. An exemplary separation of model proteins by cation exchange chromatography followed by desalting is shown. This approach can easily be adapted to various protein purification protocols.
Chromatogram of the cation exchange method.
| Method | Two step purification |
|---|---|
| Mode | IEC |
| Substances | Cytochrome C, Ribonuklease A |
| Key words | FPLC, IEC, protein purification, Cytochrome C, Ribonuklease A, AZURA® Bio Lab system |
| CAS number | Cytochrome C: CAS 9007-43-6; Ribonuklease A: CAS 9001-99-4 |
| Version | Version 1 / 12/2020 |
AZURA® Pump P 6.1L (LPG), with 10 ml pump head (ceramic), degasser and mixer (250 µl)
Article No.: APH64EB
Show product
Left module: UV detector UVD 2.1S - EDA03XA Middle module: Valve drive VU 4.1 - EWA04 Right module: Valve drive VU 4.1 - EWA04
Article No.: AY00001
Show product
Assistant module UVD 2.1S without flow cell
Article No.: EDA03XA
Show product
Assistant module VU 4.1 for valves V 4.1
Article No.: EWA04
Show product
3 mm path length, 2 µl, 1/16", 30 bar, biocompatible, classical KNAUER flow cell
Article No.: A4045
Show product
Multiposition valve, 8 Port, PEEK, PEEK, 1/16'', 240 bar, 0.75 mm bore size
Article No.: AVS34CE
Show product
Column Selection Valve VICI for 5 columns, 1 bypass, reverse flow, 12 ports, 1/16“, 50 bar
Article No.: AVZ52CE
Show product
VU 4.1 valve drive for V 4.1 valves
Article No.: AWA01XA
Show product
Fraction collector Foxy® R1 for 1/16“ or 1/8“ tubing
Article No.: A59100
Show product
Basic License for one system
Article No.: A2650
Show product
Prepacked 1 ml, 100 µm, 3 bar
Article No.: 010X15QSPZ
Show product
Prepacked 5 ml, 20 - 50 µm, 3 bar
Article No.: 040X460SPZ
Show product
for up to 50 mL/min
Article No.: SYS875413122
Show productWe are happy to help you