Appendix C Cleanup Of Proteins; Overview - LC Packings SWITCHOS II User Manual

APPENDIX C
C.1
This appendix describes a common application of the Switchos II™ Advanced
Micro Switching Unit and is provided to give the user an understanding of the
power and flexibility of the system. In this application the two 10-port switching
valves are used for on-line sample clean-up, digestion of proteins and peptide
separation.
An ion-exchange column and protein digestion column are connected to valve A,
and a reversed phase trap column and the capillary/nano separation column are
connected to valve B as shown in FIGURE C-1.
The following steps are involved in the process:
a) Proteins are injected and trapped onto the ion-exchange cartridge.
b) Valve A is placed in position 10-1 and non-charged compounds, e.g.
detergents, are flushed to waste.
c) Valve A is switched back into position 1-2. The ion-exchange column is now
in series with a protein digestion column and the reversed phase trap column,
mounted on valve B (position 10-1). Proteins are eluted by injection of a salt
solution. The peptides are concentrated onto the reversed phase trap column.
d) Valve B is switched back from position 10-1 the 1-2 to place the trap column
and separation column in series. The gradient is started to elute the peptides
from the trap column. The Peptides are separated on the capillary/nano
column.
User's Manual Switchos II
Cleanup of Proteins

Overview

D850R3
C-1