Tips for increasing the life of SEC columns - WKB121506

ENVIRONMENT

• Column life
• BEH
• Size exclusion chromatography (SEC)
• XBridge
• ACQUITY
• BioResolve SEC mAb
• BioSuite SEC
• ACQUITY Premier Protein SEC
• care and use
• tips

ANSWER

The SEC column lifetime is affected by particulates and microbial growth due to buffers at biological pH.

ADDITIONAL INFORMATION

For the shared system:

1) SEC mobile phase containers are not recommended to use mobile phase filters because they are a major source of microbial contamination in SEC columns.

2) Institute the following procedure to prepare the lines prior to hooking up the SEC column (taken from page 29 of the optimization guide (link below):

3) Needle washes and seal washes older than two weeks should be reprepared when using the SEC columns on the system.

For Mobile phase preparation:

1) The water used must be high-purity water (18.2 megaohm) when preparing the SEC mobile phases. If bottled water is used, it must be opened the day of use.

2) All buffered mobile phase should be passed through a sterile 0.22-µm filter. Avoid sintered glass filters; they leach silicates into the mobile phase, which further fouls the SEC column bed and lowers column lifetime.

3) If mobile phase contains salt less than 150mM, it must be changed out to fresh buffer, in a fresh container, every three days. 

It will promote the growth of microbes if allowed to sit longer, and if the bottle is topped off with fresh mobile phase. 

4) Always change the bottles/containers when replacing the mobile phase.

5) All mobile phase containers should be visually inspected daily for microbial growth and/or particulates. Microbial growth may form a film on the bottle surface, which can be observed by swirling the bottle. 

See also the “Trust my Science” eight-minute video for further tips on microbial growth in mobile phases.

https://www.youtube.com/watch?v=5b_WYgDVtOA

For additional information, see the Guide to SEC of mAb's, aggregates, monomers and fragements and How should I clean an ACQUITY BEH SEC column or XBridge BEH SEC column if I suspect protein has precipitated on the column?