Large Scale Albumin Depletion - (Feb/08/2010 )
I am purifying an IgG fraction from whole serum and am dissatisfied with the albumin depletion methods we use. They are expensive and time consuming as well as being ineffective at the best of times. We currently use Sephadex to remove a large portion and then species-specific columns of bound Ab to clean up the remainder. The Sephadex removes a large chunk of the albumin but at the volumes we do (up to 2L) there still can be a required 50ml~100ml pass over the gravity column. This can only be done about two or three times a day with attention needing to be payed to it as well. All in all, it can add up to a month long process quickly.
I am looking for a more efficient way of depleting albumin in our samples while maintaining the IgG fraction. I would like it to also be more cost effective than our current procedure. I recently purchased some Cibacrom Blue F3GA as that seemed like a promising idea but have been unsuccessful in finding a protocol to bind it to desalting gel beads or any other immobilization technique.
Any large scale Albumin depletion techniques would be appreciated, or a resource with Cib Blue F3GA immobilization techniques.
you can purchase immobilized Cib Blue F3GA from ge healthcare.
but it is not the best way to deplete albumin. it will also bind other proteins in which you may be interested.
beckman-coulter markets a kit which uses immobilized igy to deplete albumin but i don't know if it is available for the scale that you require.