Hello there folks,
I'm about to start working in a Agilent system using a MAbPac SEC-1 column. As the system was not functioning for a while I need to thoroughly clean it before actually doing sample analysis.
The column manufacturer guideline's describe a cleaning routine, as below, which I believe is a common practice.
1. Wash the column with 100% D.I. water at 200 µL /min for 30 minutes.
2. Wash the column with 80% methanol at 200 µL /min for 30 minutes.
3. Wash the column with 0.1 M ammonium acetate pH 5.0 buffer at 200 µL /min for 30 minutes.
4. Wash the column with the mobile phase at 200 µL /min for at least 30 minutes.
However, I do not posses ammonium acetate right now, only ammonium formate. I know they differ in pKa and buffering ranges (max pH 4.8) , so, could you guys share if anyone used it for cleaning purposes or if would not be suitable?
Thanks!
MAbPac SEC-1 is a size exclusion chromatography column used for the analysis of monoclonal antibodies (MAbs) and other large biomolecules. Here is a general cleaning routine for the MAbPac SEC-1 column:
Materials:
Distilled water
Methanol
Sodium hydroxide (NaOH)
Hydrochloric acid (HCl)
Buffer solution (if needed)
Column storage solution (recommended)
Procedure:
After completing your chromatography run, flush the column with at least 5 column volumes of distilled water to remove any buffer salts.
If needed, wash the column with a methanol-water mixture (e.g., 50:50) to remove any nonpolar contaminants.
Equilibrate the column with 20 mM NaOH for 30-60 minutes to remove any protein residues.
Rinse the column with 20 mM HCl to neutralize the NaOH and remove any remaining protein residues.
If needed, wash the column with the appropriate buffer solution to remove any remaining sample residues.
Flush the column with at least 5 column volumes of distilled water to remove any residual buffer salts.
Store the column in the recommended storage solution to prevent microbial growth and drying out.
Note: Always follow the manufacturer's instructions for cleaning and storage of the MAbPac SEC-1 column. Avoid using harsh chemicals or solvents that may damage the column material. Additionally, if you are planning to reuse the column, it is important to check the efficiency of the column using a standard sample before reusing it for analysis.
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