Hi everyone,
I'll work on zirconium-89 radiolabeled antibodies and would like to have some help for conjugate antibodies purification. I'm thinking about using PD10 column, is it a good idea ?
The PD10 column is pre-packed gel filtration column for "desalting," allowing the removal of low molecular weight substances from macromolecules, like antibodies. The macromolecules pass through easily and elute in the void volume, whereas low molecular weight substances remain in the column, which can be discarded if desired. The PD10 column can be used in gravity elution mode or in centrifugation mode. The advantage of the centrifuge column method is that the macromolecule is not diluted by the procedure.
If your labeling reagent is of a low molecular weight such that it separates from the antibody, you can use the PD10 to remove the excess reagent.
Depending on the volume of your sample, there are several flavors of Sephadex G-25 filled columns, of which PD-10 is one. NAP-5 is another (smaller) one. Check also spin columns for PCR purification and the contents of purification kits for labeled antibodies/antibody labeling kits. There are some on the market for fluorescein or Alexa dye labeling.
Read also on the various Sephadex resins at the Cytiva website.
In general, Sephadex is re-usable. Just flush with 10 column volumes and keep it on PBSN3 or 20% EtOH to prevent microbial growth.
I appreciate your help.
We'll be working on pembrolizumab for a start so we'll work on small quantities.
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